Fallypride IND Application
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Fallypride IND Application
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Fallypride IND Application
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B. NID...
Fallypride IND Application
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Fallypride IND Application
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Fallypride IND Application
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Fallypride IND Application
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Fallypride IND Application
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Table 4: [18F]Fallypride Synthesis Process Results



             [F-18]   Yield (% - specific              Kryptofix
   ...
3/20/07     240       24.32      7.92       100.00    <50 µg/mL    5.50   <0.125 EU/mL No Growth

 3/28/07     354       2...
C.6. Manufacturing Steps.

       Cyclotron production of [18F]fluoride in the Advance Cyclotron Systems TR19.

    The nu...
The radioactive peak of [18F]Fallypride, with a retention time of RT ≈ 13min
corresponding to the “cold” Fallypride standa...
3.5) and CH3CN (35: 65 v/v) at a flow rate of 2 mL/min is used to elute the
              radioligand, which has a retenti...
3)      Total radioactivity assay – every production batch before the product vial is
    released for injection

   4)   ...
[F-18]Fallypride

                                          Lot No.:____________
                 Sterile, nonpyrogenic so...
were in close agreement, including the use of the cerebellum as a reference tissue
instead of the arterial input function....
1) Acute Toxicity in Rats. The maximal dose in rats (600 µg/kg) caused mild sedation, no deaths, and no
abnormalities on p...
Tabulation of Radiation Doses to Subjects
Radioactive Material or Procedure: [18F]fallypride
Administered Quantity or View...
Testes                                                                         0.13 [0.52]

Thymus                        ...
XI.     References.

Adler LE, Hoffer LJ, Griffith J, Waldo MC, Freedman R (1992): Normalization by nicotine of deficient
...
Gatley SJ, Ding YS, Brady D, Gifford AN, Dewey SL, Carroll FI, Fowler JS , Volkow ND (1998): In
        vitro and ex vivo ...
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  1. 1. Fallypride IND Application June 22, 2007 Investigator-Sponsored IND Application: PET Imaging of Brain Dopamine Receptors with [18F]N-allyl-5-fluoropropylepidepride Injection Investigators Edythe D. London, Ph.D. Professor of Psychiatry and Biobehavioral Sciences; and Professor of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California Los Angeles Buyean Lee, Ph.D. Assitant Researcher, Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California Los Angeles Mark Mandelkern, M.D., Ph.D. Clinical Professor of Radiological Sciences, David Geffen School of Medicine, University of California Los Angeles; and Director of Positron Emission Tomography, Veterans Administration Greater Los Angeles Health System Judah Farahi, Ph.D. Chief Chemist, ??the name of the lab and department??Veterans Administration Greater Los Angeles Health System Investigator to whom correspondence should be addressed: Edythe D. London, Ph.D. UCLA Semel Institute for Neuroscience and Human Behavior 760 Westwood Plaza Room C8-532 Los Angeles, CA 90024 Mail Code: 175919 Page 1 of 37
  2. 2. Fallypride IND Application June 22, 2007 CONTENTS Form FDA 1571, Investigational New Drug Applications i Form FDA 1572, Statement of Investigator ii Biosketches iii I. Introductory Statement of Purpose and General Plan 5 II. Initial Study Protocol: 6 A. Purpose 6 B. Background 6 C. Subject Selection 9 C.1. Inclusion Criteria 9 C.2. Exclusion Criteria 9 D. Research Procedures 10 E. Data Analysis 12 III. Chemistry, Preparation, and Control 13 A. Chemistry and Manufacturing Introduction 13 B. Drug Substance 13 B.1. A Description of the Drug Substance and Drug Product 13 B.2. Characterization of the Drug Substance 14 C. Drug Product 14 C.1. Composition of the Individual Dose of the Drug Product 15 C.2. Batch Formula Quantity 15 C.3. List of Components of the Drug Product by Name 16 C.4. Preliminary Production Data 16 C.5. Facilities for Preparation and Quality Control 18 C.6. Manufacturing Steps 18 C.7. Quality Control 19 IV. Pharmacology and Toxicology 25 V. Dosimetry 31 IX. Human Experience 34 X. Environmental Assessment 34 XI. Case Report Form 35 XII. References 38 XIII. Appendices A. Human Study Protocol Page 2 of 37
  3. 3. Fallypride IND Application June 22, 2007 B. NIDA PET Radiochemistry Facility Floor Plan C. Synthesis of 2-[18F]F-A-85380 Injection Batch Record D. Bacterial Reverse Mutation Assay E. Convulsive Dose ED50 Dose-Response Curves F. Acute, Two-Day Intravenous Toxicity Study of 2-Fluoro- 3-(2(S)- azetidinylmethoxy)pyridine (2-F-A-85380) in the Mouse G. Expanded, Acute, 14-Day Intravenous Toxicity Study of 2-Fluoro-3- (2(S)-azetidinylmethoxy)pyridine (2-F-A-85380) in the Mouse H. Acute Cardiovascular, Locomotor, and Temperature Effects of 2- Fluoro-3-(2(S)-azetidinylmethoxy)pyridine (2-F-A-85380) in the Unanesthetized Rat I. Acute Effects of 2-Fluoro-3-(2(S)-azetidinylmethoxy)pyridine (2-F-A-85380) on the Electrocardiogram of the Unanesthetized Rat J. Acute and Prolonged Effects of 2-F-A-85380 on the Hematology and Blood Chemistry of the Rhesus Monkey Page 3 of 37
  4. 4. Fallypride IND Application June 22, 2007 I. Introductory Statement of Purpose and General Plan Fallypride, N-allyl-5-fluoropropylepidepride, is a selective, high affinity D2-like dopamine receptor antagonist with KD of 30 pM (Christian et al, 2000; Kessler et al, 1993). The utility of this compound is implemented in Positron Emission Tomography (PET) imaging for the purposes of the current protocol entitled Early Methamphetamine Abstinence: fMRI and Cognition, an ongoing human research project at the University of California Los Angeles, Laboratory of Molecular Neuroimaging. Individuals who abuse methamphetamine (MA) exhibit deficits in markers for dopaminergic neurotransmission including D2/D3 dopamine receptors. For example, previous brain imaging studies with PET have reported that MA abusers have lower level of D2/D3 dopamine receptors than healthy control subjects in the striatum of the brain; however, assessment of extrastriatal dopamine receptors in MA abusers has not been reported. Since it is now possible to assay extrastriatal D2/D3 dopamine receptors in human subjects noninvasively with PET, using [18F]fallypride, we use this radiopharmaceutical in assessment of D2/D3 dopamine receptors in the brain of human subject under authorization of the UCLA Institutional Review Board and the Radioactive Drug Research Committee. The range of radiation exposure from [18F]fallypride given to research subjects in this study is between 606-1213 mRem [BUYEAN – is this the EDE?, yes]. This range is well below the exposure limit sanctioned by FDA, 21 CFR 361.1, and is associated with no known toxic effects. This proposal aims to accomplish the following: 1) This IND application is to secure approval for the use of [18F]fallypride, as the Drug Product ([18F]fallypride Injection), for further research purposes. There is no intention to use the Drug Product for clinical studies involving diagnosis or treatment. The following investigators are collaborating to accomplish this goal: Edythe D. London, Ph.D. Professor of Psychiatry & Behavioral Sciences Professor of Molecular & Medical Pharmacology Buyean Lee, Ph.D. Assistant Researcher Mark Mandelkern, M.D., Ph.D. Clinical Professor of Radiological Sciences Judah Farahi, Ph.D. Chief Chemist,???? [Please change all the titles to match the front page.] II. Human Study Protocol: Early Methamphetamine Abstinence: fMRI and Cognition A. Purpose This project identifies the brain regions and pathways that show dysfunction contributing to the problems of Methamphetamine (MA) abusers in performing mental tasks. We use functional magnetic resonance imaging (fMRI), a noninvasive brain imaging procedure, to study brain function while research participants perform tests of memory and concentration. We also perform positron emission tomography (PET), another noninvasive imagining procedure, to relate the integrity of dopamine receptors in brain to performance on tests of memory and concentration. We compare brain activity in regular users of MA with activity in a comparison group of participants who do not use MA. B. Background Page 4 of 37
  5. 5. Fallypride IND Application June 22, 2007 No effective pharmacological treatments for MA dependence are known. The major therapeutic strategies are psychosocial approaches that depend upon the client’s ability to process and remember information.1 The cognitive impairments suffered by MA abusers, therefore, can interfere with treatment. MA users exhibit deficits when performing tasks that require cognitive control, including tests of working memory and selective attention.2,3 Knowledge of the cognitive limitations of MA abusers can help design treatments that would consider these problems. This work builds on knowledge of the circuitry important to cognitive functions that are impaired in MA abusers, using fMRI to help define the biological determinants of the deficits. Neuroimaging techniques can help delineate function during memory and attention processing.4 As MA abusers show deficits in these cognitive functions, we anticipate that the normal pattern and intensity of activation may not be observed in MA abusers. Our data from an ongoing study (protocol # 98-09-018-11) show that MA abusers in early abstinence (4-7 days) have deficits in these cognitive functions and abnormalities in the metabolism of brain regions that mediate them.5-12 Although brain metabolism increased in a subset of participants who remained abstinent from MA for an additional month, dopaminergic terminal field areas showed less of an increase than frontal cortical areas. We, therefore, anticipate that our fMRI study will reveal abnormalities in activation during the first week of abstinence, and that activation will change with continued abstinence. As aforementioned, it is well-established that MA abusers exhibit deficits in markers for dopaminergic neurotransmission. Positron emission tomography (PET) scanning has demonstrated deficits in dopamine transporter (DAT) molecules in the striatum, and also in the orbitofrontal, anterior cingulate, and insular cortices and amygdala. PET studies have also identified reduced availability of D2-like dopamine receptors in the striatum; however, assessment of extrastriatal dopamine receptors in MA users has not been reported. Since it is now possible to assay extrastriatal D2/D3 dopamine receptors in human subjects noninvasively with PET, using [18F]fallypride13, we use this radiopharmaceutical in assessment of cerebral D2/D3 dopamine receptors, including cortical and amygdalar measures as a function of MA abstinence and in relation to cognitive activation, which we measure with fMRI. In rodents and humans 14-20, methamphetamine treatment mimics many of the behavioral effects of bipolar disorder. Many manic features are observed during the activation phase of methamphetamine intoxication including elevated mood, increased energy, hyperlocomotion, perseverations, hypersexuality, decreased appetite, and hyposomnia. During the withdrawal phase, on the other hand, depressive features are observed, including low mood, low energy, decreased locomotion, passivity, and hedonia, and hypersomnia. Nonetheless, we are aware of only one published neuroimaging study 20 that explores relationships between methamphetamine and bipolar disorder in humans. Recently, a proton magnetic resonance spectroscopy (1H MRS) investigation conducted by our UCLA collaborators and colleagues detected above-normal levels of glutamate and creatine+phosphocreatine in the mesial prefrontal cortex in adult bipolar depressed patients. The relatively large volume sampled comprised a mixture of bilateral perigenual anterior cingulate, anterior middle cingulate, and superior frontal cortices. Measuring Brain Function with fMRI. We pair fMRI with cognitive/behavioral tasks to test brain function. The approach depends on the fact that changes in the magnetic resonance signal accompany parallel changes in brain activity. We use the BOLD (Blood Oxygenation Level Dependent) approach,21, 22 which reflects local increases in blood flow that accompany increases in local brain function,23 and takes advantage of the different magnetic susceptibilities (χ) of the oxygenated and deoxygenated forms of hemoglobin. The greater χ of deoxyhemoglobin produces a local perturbation of the magnetic field and, therefore, a decrease in the amplitude of the MR signal.24 Because the extraction fraction of oxygen across the capillaries decreases in Page 5 of 37
  6. 6. Fallypride IND Application June 22, 2007 the face of increased blood flow, the oxygen concentration increases in the post-capillary venules.25 The post-capillary oxyhemoglobin increases and so, therefore, does the MR signal.21 1.5T MRI scan sessions. Participants have three scan sessions in the 1.5T MRI scanner (2 sessions in Phase 1, 1 session in Phase 2). In addition to acquiring a structural scan, proton magnetic resonance spectroscopy (1H MRS) is performed. 1H MRS is a noninvasive technique which allows identification of molecular structures, including metabolite abnormalities, and their amounts by detecting energy exchange between external magnetic fields and Hydrogen nuclei in molecules. Methamphetamine-dependent individuals exhibit similarities to bipolar depressed patients and 1H MRS is able to assess abnormalities in metabolite concentration (glutamate and creatine+phosphocreatine). C. Subject Selection This ongoing protocol projects an enrollment of 250 subjects: 160 MA abusers and 90 control subjects (with no current or prior illicit drug dependence). Based on experience the attrition rate and the difficulty of finding participants who meet inclusion criteria, we anticipate enrolling 250 subjects in order to have 54 completers (27/group). We expect PET and fMRI data from ~10% of the completers (6 subjects) to be unusable due to technical problems (e.g., motion artifact), and data from the other 48 completers (24/group) to be usable. C.1. Inclusion Criteria: (1) Age and Gender: 21 to 50 years of age, both genders. (2) Handedness : Right-handedness (score >20 on a modified Edinburgh Handedness Test).45 C.2. Exclusion Criteria: (1) Psychiatric disease: Current or lifetime history of an Axis I diagnosis (according to DSM-IV criteria), except for the diagnosis of MA dependence (MA abuser group only) and the diagnosis of nicotine dependence (both groups). The diagnoses of post-traumatic stress disorder and childhood ADHD will be exclusionary in both groups. (2) History of Drug Abuse: Potential participants with dependence on any drug (other than nicotine for controls, or, for the MA group, MA) will be excluded. (3) Medications: Evidence of taking psychotropic medications (other than MA for the MA abuser group and nicotine for both groups) will be exclusionary for both groups (4) CNS Disease: Including structural brain abnormalities (e.g., neoplasms, subarachnoid cysts), cerebrovascular disease, infectious disease (e.g., abscess), history of other neurological disease, including stroke or head trauma (defined as loss of consciousness > 5 min or requiring hospitalization). (5) Cardiovascular Disease: Advanced coronary artery disease, endocarditis or other cardiac disease likely to result in cerebral embolism. (6) Pulmonary Disease: Significant obstructive pulmonary disease or tuberculosis. (7) Systemic Disease: Endocrinopathies, renal or hepatic failure, or autoimmune disease involving the CNS. Page 6 of 37
  7. 7. Fallypride IND Application June 22, 2007 (8) Claustrophobia: Subjects will be questioned about possible discomfort with being in an enclosed space (e.g., MRI scanner). Those who report such problems will be excluded. (9) Pregnancy: Pregnancy, possible pregnancy, or failure to indicate the use of reliable birth control precautions. We will administer a pregnancy test to each female participant before starting each of the 2 phases of the study. Subjects must agree to use an effective form of birth control between participation in Phase I and Phase II of the study. The only acceptable methods of birth control are: 1) oral contraceptive; 2) Depo-Provera injection; 3) intrauterine device; 4) diaphragm with contraceptive jelly; 5) condom with contraceptive jelly, foam, or sponge; 6) tubal ligation or hysterectomy; 7) abstinence from heterosexual intercourse. (10) HIV: Subjects who are HIV-seropositive will be excluded because HIV infection and the development of AIDS produce alterations in brain activity that are dependent on the stage of AIDS46-48. Such effects on brain activity would confound the results of this study. (11) We will exclude any subject whose body contains a ferromagnetic implanted device that might produce a safety hazard during fMRI. We will use as guidelines, the manual, Magnetic Resonance: Bioeffects, Safety, and Patient Management49 supplemented by the current information published on the International MR Safety Web Site: http://kanal.arad.upmc.edu/mrsafety.html. (12) Vision: Colorblindness, which could affect performance on the Stroop Task. (13) Language: Lack of fluency in English will be an exclusion criterion because it would interfere with performance of psychological tests and filling out questionnaires used in the study. The consent form, all questionnaires and instructions will be given in English. (14) Neurological: Neurological that is not within normal limits as determined by an authorized study physician and as indicated in self-report. (15)Radiation Exposure: Participated in any other research study involving exposure to ionizing radiation in the past year if the total cumulative exposure from the past research studies and the current research study would exceed the limits described by the FDA in 21 CFR 361.1. Specifically the total cumulated dose to the whole body, active blood-forming organs, lens of the eye, and gonads must remain below 5 rems and the cumulated dose to all other organs must remain below 15 rems. Potential subjects who have had exposure to ionizing radiation in the past year cannot be allowed to participate if we are unable to obtain proper documentation quantifying the amount of past exposure. (16)Miscellaneous: hematocrit <32, prostatic hypertrophy or chronic inflammation. (17) Control Group: Must smoke at least 15 cigarettes per day for the past 2 years (as indicated by self-report) D. Research Procedures After the initial telephone screening, participants will visit our laboratory to provide informed consent. Having given consent, the participants will undergo comprehensive evaluation to determine final eligibility status. The evaluation will include a psychiatric diagnostic interview according to the Structured Clinical Inventory for DSM-IV (SCID-IV),51 a medical history and physical examination, and blood tests [including a complete Page 7 of 37
  8. 8. Fallypride IND Application June 22, 2007 chemistry panel (5 ml), hepatic panel (3 ml), and HIV test (5 ml)]. Urine samples will be taken for drug screening, and to test for pregnancy (female subjects). Subjects will be interviewed about their prior and current drug use. Those providing evidence of a past diagnosis of drug dependence other than relevant MA abuse (or nicotine dependence), or other significant current drug use within 2 weeks of entering the study based on self-report, SCID, and/or urine toxicology results, will be excluded, except in cases of light alcohol (≤ 10 drinks of liquor or the equivalent of beer or wine per week) or occasional cannabis (≤ 1 joint per week) use and tobacco dependence, which will not be exclusion criteria. Subjects will be assigned to one of two groups based on history of drug abuse (MA abuser or control) and will be matched on gender and age (within a 5-yr range), ethnicity, and mother's level of education. MA abusers will be accepted into the study only after successful completion of the following: informed consent, completion of an interview and questionnaires pertaining to drug use history, collection of a urine sample that is positive for MA and negative for other illicit drugs of abuse, and collection of a saliva test that is negative for alcohol. Control subjects will also be interviewed, will answer questionnaires about drug use history, and will provide urine and saliva samples for drug screening. Subjects in the MA group will be admitted to the General Clinical Research Center (CRC) and will participate on a residential basis. During their hospital stay, subjects will have access to phones and recreation facilities, such as TV, games and music, and will participate in occupational therapy. While participants are in the hospital, their health will be monitored closely by a team of doctors and nurses. The nurses will daily measure weight and caloric intake as part of our study of changes, including remarkable weight gain associated with early abstinence from MA in dependent research participants. Control subjects will participate on a non-residential basis. Measures Obtained in Addition to fMRI Testing (after screening is complete) Urine and Blood Tests Urine samples will be collected daily while MA abusers are residing at our facilities and the samples will be used for random (twice each week) screening for cocaine, amphetamines, opiates, cannabinoids, and benzodiazepines. A saliva test (for alcohol) and urine screen will be administered to control subjects on testing days only. The MA abusers will have the corresponding urine and saliva tests on fMRI and cognitive testing days. Any subject in the MA group who tests positive for illicit drugs while residing as an inpatient at UCLA will be discharged. S/he will receive compensation for participation up to that point. Control subjects who test positive for illicit drugs of abuse other than marijuana will be excluded from further study, but will receive compensation for participation up to that point. If their urine test indicates recent use of marijuana, they can return for re-testing, as once weekly use of marijuana is not an exclusion criterion. We take up to two blood samples (5 ml. each) from control subjects and three blood samples (5 ml. each) for assay of cortisol, prolactin, and adrenocorticoptropic hormone. Measurements of these stress hormones reflect hypothalamic-pituitary-adrenal (HPA) axis functioning. Research has shown that abnormalities of the HPA axis are related to predisposition to substance abuse; however, data related to the predisposition to MA Page 8 of 37
  9. 9. Fallypride IND Application June 22, 2007 abuse in particular have not yet been reported. Our study will address the question of the level of function in the HPA in MA users. Participants also have blood samples taken for HIV testing, Subjects will be asked to sign an additional consent form at the General Clinical Research Center for HIV testing. Study staff will explain the HIV test and the implication of a positive HIV test result. Staff also inform subjects that if he/she is found to be HIV-positive, he/she be contacted by the study physician who arrange a meeting over the phone or in person to discuss his/her HIV status. The study physician inform HIV-positive participants of the implications of their HIV-positive status as well as refer subjects to further information and appropriate services. Study staff inform subjects with negative HIV-test results of their HIV status. A blood sample (10 ml) be taken from each subject for genetic testing (at UCLA, in collaboration with Drs. Terry Ritchie and Ernest Noble). Several genetic polymorphisms related to neurotransmitter function (e.g., alleles for dopamine receptors, transporters) have been related to substance abuse. Others (e.g., alleles for catechol-O- methyltransferase, COMT) have been associated with variation in hemodynamic responses to cognitive activation. We relate measures of impulsivity, drug history and cognitive activation with some of these markers. DNA be extracted and stored at UCLA. We plan to assay first for markers related to dopamine neurotransmission (dopamine receptors, dopamine transporter, COMT). We may assay for other markers as relevant literature dictates. Questionnaires, Including Those Related to Drug Use All subjects: Study Admission Form (SAF). A 5-page questionnaire developed at the Los Angeles Addiction Research Consortium. Drug Use Survey, a 2-page questionnaire developed at the Intramural Research Program, NIDA. Neurological History Questionnaire, a 2-page questionnaire administered at baseline, designed to screen for brain dysfunction. Smoker’s Profile, a 2-page questionnaire administered at baseline, designed collect cigarette smoking demographics. Fagerstrom Tolerance Questionnaire, a 1-page questionnaire administered at baseline, designed to determine nicotine dependence. MA Abusers only: Visual Analog Scales (VAS) of MA (and other drug) craving and withdrawal symptoms are administered at baseline and on each day of study participation, and will take approximately 5 min. Addiction Severity Index (ASI).52 This questionnaire takes approximately 45-60 min to complete. It will provide demographic information and composite scores on the level of functioning within each psychosocial area. Additional Questionnaires Related to Psychiatric Diagnosis and Mood/Affect: Wender Utah Rating Scale for ADHD.53 This measure will be used to diagnose probable childhood Attention-Deficit Hyperactivity Disorder (ADHD). A cutoff score of 46 will be used on a subset of items specific as a diagnostic criterion for history of ADHD. The questionnaire takes approximately 10 min. Page 9 of 37
  10. 10. Fallypride IND Application June 22, 2007 ADHD Questionnaire. This questionnaire assesses symptoms of ADHD over two time periods: the present and during childhood/adolescence. The questionnaire will take approximately 3 min. Beck Depression Inventory (BDI).54 This 21-item questionnaire evaluates symptoms of depression. This self-report instrument will be administered at baseline and on each day of study participation for the MA abusers, and during baseline measurement only for the control subjects. It takes approximately 5 min. Symptom Check List (SCL-90-R).55 This is a self-report assessment of the degree of psychological distress and prevalence of symptoms endorsed within each of nine psychological state-related dimensions and three global indices of distress. Changes in scores will be compared to drug use group and status of drug withdrawal. The self-report will be administered at baseline and on each day of study participation for the MA abusers, and only once (at baseline measurement) for the control subjects. It takes approximately 10 min. State-Trait Anxiety Inventory (STAI) Self-Evaluation Questionnaire forms Y-1 and Y-2.56 The STAI is a self-report questionnaire that assesses the subject’s state and trait anxiety. The STAI will be administered to both groups of subjects on each day of fMRI scanning. Brief Symptom Inventory.57 This questionnaire assesses symptoms of mental disorder. The questionnaire assesses the clinical dimensions of 1) Somatization 2) Obsessive- Compulsiveness, 3) Interpersonal Sensitivity, 4) Depression, 5) Anxiety, 6) Hostility, 7) Phobic Anxiety 8) Paranoid Ideation, and 9) Psychoticism. A global severity measure is also obtained. The self-report will be administered at baseline and on each day of study participation for the MA abusers, and only once (at baseline measurement) for the control subjects. It takes approximately 5 min. Hamilton Anxiety Scale.58 This 14-item questionnaire evaluates symptoms of anxiety. This observer rating will be administered at baseline and on each day of study participation for the MA abusers, and only once (at baseline measurement) for the control subjects. It will take approximately 5 min. Hamilton Depression Scale.59 This 24-item questionnaire (observer rating) evaluates symptoms of depression. It will be administered at baseline and on each day of study participation for the MA abusers, and only once (baseline measurement) for control subjects. It takes approximately 5 min. Barratt Impulsiveness Scale (BIS)60This 30 item self-reported questionnaire is a research tool designed to measure impulsiveness. Each item is scored from 0-4 with 4 indicating the most impulsive. The higher the sum, the higher the level of impulsiveness. It will be administered once, at baseline and takes approximately 5 min. South Oaks Gambling Screen (SOGS)61The SOGS is a 20-item questionnaire that queries the type of gambling behavior, largest amount gambled in a single day, parental history of problematic gambling and the largest amount gambled in a single day, and the individual’s opinion about whether his/her gambling is problematic. The SOGS is a useful research and clinical diagnostic tool in determining the lifetime prevalence of pathological gambling. It will be administered at baseline and takes approximately 10 min. URICA (University of Rhode Island Change Assessment).62The URICA is a 32-item inventory designed to assess an individual's stage of change located along a theorized Page 10 of 37
  11. 11. Fallypride IND Application June 22, 2007 continuum of change. Four 8-item subscales correspond to the each of four stages of change: Precontemplation, Contemplation, Action, and Maintenance. Items rated on a 5- point Likert scale describe how an individual might think or feel when starting therapy. A score derived from each of the subscales determines an individual's location in a particular stage of change. It will be measured at baseline and again at the end of the study. It takes 5 min. Phases of Study The study is conducted in two phases. During Phase I, we will test MA abusers at 4-7 days of abstinence from the drug. In Phase II, we will assess the effects of continued abstinence for another 4-5 wk. Control subjects will participate at corresponding times. Each phase will include a neurocognitive test battery, structural MRI, proton MR spectroscopy (1H MRS), PET, and cognitive activation with fMRI. The primary dependent measures will be accuracy and reaction times on cognitive tests and task- induced changes in fMRI signal intensity. There will be three separate fMRI sessions (A, B, and C) in each phase, with the order counterbalanced across subjects. Each session will last about 60 min, minimizing fatigue and the chance of artifact due to motion. The 3 test sessions may be on the same day, with a break of 30 min; but the participant may choose to split the sessions between days. However, the sessions will be no more than 48 h apart. In Session A, the primary fMRI measures will be taken during performance of the N-Back Task and control measures (visual stimuli, motor task). The control measures are included to test whether possible differences between groups reflect nonspecific effects of chronic MA on the dynamic range of BOLD rather than specific effects related directly to the cognitive functions under study. In Session B, the primary fMRI measures will be taken during performance of the Stroop Task and control measures (e.g., visual stimuli, motor task). In session C, the primary fMRI measures will be taken during performance of the Stop-Signal Task and Probabilistic Reversal Learning Tasks and control measures. Sessions will be counterbalanced across all subjects. For all subjects who complete Phase II, the session order will be the same as the order in Phase I. 1.5T scan session Participants undergo three MRI scans on the 1.5T scanner at the Ahmanson-Lovelace Brain Mapping Center. Subjects will have two 1.5T scan sessions in Phase 1 for including a structural scan MRIand proton magnetic resonance spectroscopy (1H MRS). Session 1 in Phase 1 is about 2 hours, Session 2 in Phase 1 is 30 min and Session 3 in Phase 2 is 30 min. The procedures for the 1.5T scan session will be the same as that of the fMRI scan: subjects will lie still on the scan bed and be able to watch a movie while performing the scan. Participants undergo one PET scan in each phase, receiving one dose of [18F]fallypride in each scan. Each PET scan session will take about 3 hours for positioning, injection of [18F]fallypride and the dynamic scanning sequence. The 1st set of fMRI tests (3 sessions) and PET scan take place after 4-7 days of abstinence from illicit drugs of abuse (initiated upon enrollment in the study). We accompany subjects to the Ahmanson-Lovelace Brain Mapping Building for scanning. After each session, subjects in the MA abuser group will return to the GCRC in-patient facilities, Page 11 of 37
  12. 12. Fallypride IND Application June 22, 2007 where they are housed and receive occupational therapy and group therapy (offered once weekly) until they complete their participation. Control subjects are discharged after each scanning session (unless the sessions are on the same day) and are scheduled for the next session. After completion of both fMRI sessions, PET scans, and cognitive tests in each of the 2 phases, all subjects are discharged. [Note: Those subjects who choose to complete only Phases I are discharged after completing the 3 fMRI sessions, PET scans, and cognitive tests in Phase I.] Ancillary Tests Cognitive Test Battery (given once in Phase I and again in Phase II, not concurrent with fMRI) Long Cognitive Battery (given once in Phase I and again in Phase II): Stroop Color Word Interference Test.63 This task demands a resolution of a conflict between competing attentional demands. It is another method of assessing focused attention with inhibition of a habitual response. It is compared with the single trial version of the Stroop task used in the magnet. Gambling Task.64 This computerized task assesses the ability to balance immediate rewards against long-term negative consequences. Wisconsin Card Sorting Task.65 This computerized task requires the subject to shift their decision from one sorting category to another and demonstrates flexibility in decision making and shifting of attentional sets. Card Sort Task (2 versions). A Card Sort task similar to that developed by Delis et al.66 provides an assessment of concept generation, concept identification, and concept execution. Backward Digit Span (2 versions).67 This task assesses the working memory of the subject utilizing digits. The subject is read increasingly long strings of digits and asked to repeat them backwards. Missing Digit Span (2 versions).68 This task assesses the working memory of the subject by testing the ability to identify missing digits in progressively longer digit strings. Sentence Span (2 versions).69 This task tests a subject’s verbal working memory by requiring the subject to remember the last word in each of several sentences that s/he reads aloud. Trail Making A.70 This task tests the subject’s attention, sequencing, psychomotor speed, and mental flexibility by connecting 25 numbers, randomly arranged on a page. Trail Making B.70 This task tests the subject’s attention, sequencing, psychomotor speed, and mental flexibility by connecting 25 alternating letters and numbers. Logical Problems (2 versions). This test consists of simple syllogisms and three term deductive reasoning problems consisting of two propositions and a question. Functional Flexibility (2 versions). This test requires the subject to write down alternate uses for an object and tests spontaneous flexibility. Shipley-Hartford Abstract Thinking.71 This task requires the subjects to continue a series of numbers, words, or letters using the beginning of the series as a guide, which is a test of fluid intelligence. Shipley-Hartford Vocabulary Test.71 This task measures the subject’s crystallized intelligence that with the Abstract Thinking task provides a short form IQ. Page 12 of 37
  13. 13. Fallypride IND Application June 22, 2007 Repeated Memory Test (RMT) (2 versions).3 This test requires the subject to recall and recognize pictures and words. The test provides a measure of source memory and intrusions. F-A-S.72 This tests the subject’s verbal fluency by evaluating the spontaneous reproduction of words within a given time. Selective Reminding. This is a memory task, which measures the subject’s ability to do associative clustering, category clustering, and subjective organizing. Discrimination Learning. This task requires the subject to identify the category as a shape, color and width, which is a measure of hypothesis testing and rule learning. Delay-Discounting Procedure. In this computerized task, participants are presented with a fixed set of 27 hypothetical choices between smaller immediate rewards and larger delayed rewards (e.g., “Would you prefer $10 today or $15 in 2 weeks?”). Visual - Verbal N-Back Task. In a separate session, MA abusers who agree will be given this 24-min computerized test, designed to further elucidate the problems that MA abusers have with working memory. Four 8-min versions of the N-Back Task are presented in a 2 (type of stimuli) x 2 (modality) design. The four tests are: visually presented words, visually presented non-verbal stimuli (brick walls), auditory words, and auditory non-verbal stimuli (bird songs). The four tests are equated for difficulty so that any differences found should be due to the stimuli or to the modality of presentation. Control participants may choose to take this 24-min test on the days that they take the cognitive battery. Tower of London: Administered Day 16 This tool features a problem-solving activity in which subjects must transfer blocks of different sizes from one tower to another. The activity is based on a game invented by the French mathematician, Edouard Lucas, in 1883. The rules are that you can only move one block at a time and that a larger block may never lie on top of a smaller one. The applet lets you set the number of blocks (the more blocks, the greater the difficulty). Raven’s Progressive Matrices: Administered Day 16. Provide an overall indication of ‘general intelligence’ and may be applied across a wide range of ability level, age group, and cultural background. The tests consist of sequences of abstract visual patterns, each with a part missing - which can be chosen from a range of alternative components. They are 'progressive' in that the pattern sets increase in difficulty throughout the test. Test administration time is usually about 40 minutes. Reward-Penalty Task (RPT): The RPT is designed to measure individuals64 sensitivity to rewards and punishment. On each trial, the individual is presented with some amount that they may win or lose on that trial. Some trials are worth more than others. The force of responses (pressure on button pad) along with changes in skin conductance provide objective indications of the emotional arousal during each trial. Electrodermal recording: Some of the tests administered in the long cognitive battery will be used in conjunction with electrodermal recording. The procedure used for recording will be those outlined by the 1981 consensus report on electrodermal measurement73. Two sintered-pellet silver chloride electrodes will be placed on the non- dominant hand of the subjects (one on the hypothenar eminence and one on the thenar eminence). An electrolyte gel will be used at both placement sites to improve stability of the contact. Electrodes will be fixed with double-sided adhesive electrode collars, and reinforced with tape. After affixing the electrodes, subjects will rest for 3 minutes to allow stabilization, and then a potentiator balance control will be used to balance the Page 13 of 37
  14. 14. Fallypride IND Application June 22, 2007 subject’s tonic (basal) conductance level to zero. An excitation current of 0.5 volts AC, 100 Hz will be used. A second 3 minute rest period will be used to measure the subject’s rate of spontaneous skin conductance responses (those greater than a predetermined threshold). Following this, subjects will perform one or more of the computerized tasks described above, while both the amplitude and frequency of skin conductance responses (changes from basal levels) are recorded. The computerized games during which skin conductance is recorded will be played in succession, then the electrodes will be removed, and then subjects will complete the remaining tasks. Pedometer: In order to provide adequate health and nutrition care for inpatients, we will utilize a pedometer to track subjects’ physical activity. The pedometer is a small, beeper- sized device that is clipped onto the inpatient participants’ clothing by the hip used to monitor the participants’ activity and caloric output. The pedometer uses an internal pendulum device to detect and record vertical motion (i.e. steps, jumps). Participants will be instructed to wear the pedometer at all times during their inpatient stay. Daily activity will be logged by research staff and monitored by the study physician. Calorie Count: In order to provide adequate health care for inpatients as well as evaluate any trends in metabolism and intake, dietary staff will assess nutrient and caloric intake for the first and fourth weeks of the inpatients’ stay. The calorie count requires no action on the participants’ behalf. Short Battery to be administered on each fMRI test day (estimated time = 30 min): The Stroop and the N-Back Tasks are complex tests, and there is some overlap in the cognitive functions they require. We will therefore administer several tests to measure the component processes of the N-back and Stroop tasks. d2 Test of Attention.74 This timed test of selective attention measures processing speed, rule compliance and quality of performance in response to the discrimination of similar visual stimuli. Go-No Go Task. (inhibition of an established response). A computerized version of this task patterned after Roser and Corballis75 was programmed for this study. Continuous Performance Test.74 This computerized test is a measure of sustained attention, in which the participant is asked to mark or ‘cancel’ specific letters or numbers as they appear. Visual Delayed Match to Sample. A computerized version of this well-known task similar to Hartman et al.76 was programmed for this study. WAIS-R Digit Symbol Sub-test.77 This task is a measure of psychomotor speed manipulation of information in working memory. To ensure that subjects are not fatigued during the Long Cognitive Battery, the battery may be administered in more than one session. However, each session will be within 72 h of the fMRI scan for each phase of the study. The entire cognitive battery will typically last about 3 h. ƒMRI and 1.5T scan MRI Procedures Participants will lie on the bed of the MRI scanner and will be given headphones and earplugs. They will also be given an MRI-compatible video system (goggles) through which they can watch movies of their choice (during preliminary scans and positioning). In some cases, a cloth tape or pads on the sides of the head Page 14 of 37
  15. 15. Fallypride IND Application June 22, 2007 may be used to limit head motion. During the fMRI procedures (typically 1.5 h, but always < 2 h), the participants will be asked to relax, perform behavioral or cognitive tasks, or provide self-reports of mood states. During the 1.5T scan session, participants can choose to view a movie or rest. The MRI equipment makes a variety of noises during scanning (e.g., repetitive tapping, beeping noises). These are all below the threshold for hearing damage, but we will use ear protectors and headphones that offer an additional 45-60 dB of attenuation, so that the noise that the subject experiences is not excessively loud. Most of the scans will require acquisition of multiple images, each lasting from 1 to 15 min. The subject will be asked to remain still during the imaging period, but can move more freely during the 2-5 min that separates the imaging series. After completing the imaging, the subject will be removed from the scanner, and we will ask about any adverse or unexpected reactions that may have taken place during scanning (none have been reported so far), and administer questionnaires related to mood states. PET Procedures The procedure for PET scans in human subjects, using [18F]fallypride for the estimation of binding potential (an indication of receptor density) have been described 13. Research participants will be prepared with 2 intravenous catheters (one in each arm) for radiopharmaceutical administration and blood sampling, respectively. Each also will be fitted with a thermoplastic facemask. They will be positioned in the scanner in a dimly lit, quiet room; and they will be instructed to lie still during the PET data acquisition. They will be free to open and shut their eyes. PET data are acquired with an EXACT HR+ PET scanner in 3-D mode. A 22-min transmission scan using 68Ge/68Ga rod sources are performed prior to administration of the radiopharmaceutical, to obtain data for measured attenuation correction. Scanning will be initiated at the start of an intravenous bolus and infusion of the radiopharmaceutical, which are followed by a saline flush. A dose of approximately 0.07 mCi/kg (maximum of 5 mCi with a maximum mass dose 1.82µg) [18F]fallypride are administered per study to each of the subjects. The first dynamic scanning sequence will take 80minutes (six frames at 30 sec, seven frames at 1 min, five frames at 2 min, four frames at 5 min, and four frames at 10 min). The subject’s facemask are marked for repositioning, and the participant are removed for a 20-min break. A second dynamic sequence of 80 minutes are acquired, yielding a total study time of approximately 3 h. Before each set of PET scans, we collect three venous blood samples (5 ml each) to determine the plasma concentration of the radiotracer and determination of metabolites. Total blood sampling during each scan will be 30 ml. We provide information on treatment programs at UCLA and other sites in the Los Angeles area for all our subjects. Information and referrals for medical treatment and other social services will also be provided for those subjects who are interested. Subjects receive monetary compensation for their participation in the study. Page 15 of 37
  16. 16. Fallypride IND Application June 22, 2007 If a subject terminates his/her participation in the study or is excluded at some point during the experiment, he/she will be compensated in accordance to the number of days they participated. Six subjects, three male and three female, will be required for Experimental Procedure 2. To achieve the goals of Experimental Procedure 2, scans will be acquired at three bed positions to put the liver, urinary bladder, and gonads within the field of view of the PET scanner. Three scanning sessions, of 1.5-, 1.8-, and 2.3-h duration (a total of 5.6 h in scanner), acquired over a 6.1-h period will include two 15-min rest breaks. The data acquired from this scanning schedule will provide the concentration of radioactivity in the target organs at 9 time points). Measurement of the electrocardiogram, heart rate, respiration rate, and blood pressure, and collection of urine samples and arterial or arterialized venous blood samples will be obtained following the schedule described in Experimental Procedure 1. E. Data Analysis Data will be spatially normalized, co-registered and corrected for motion-related signal changes using the scheme of Woods {Watson et al., 1993, REF#10253}, {Woods et al., 1992, REF#10271}, {Woods et al., 1999, REF#10404} to co-register other images with the middle image in the data set. Linear and non-linear transformations will register each image into a standard stereotactic coordinate space defined by the template image supplied in SPM2. This template conforms to definitions by the International Consortium for Brain Mapping {Mazziotta et al., 1995, REF#10099} and closely approximates the space described in the atlas of Talairach and Tournoux {Talairach and Tournoux 1988, REF#10334}. Images will be smoothed and normalized to the global signal. Analyses will be performed in two stages using SPM2 {Friston et al., 1995, REF#10407}, {Friston et al., 1995, REF#10762}, {Friston et al., 1994, REF#10760}, {Price and Friston 1996, REF#10287}. In the first stage, fMRI data of each participant will be analyzed using a fixed- effects model. Next, a single image for a given contrast will be created for each participant (e.g., in case of parametric N-Back task we will create one image based on relative differences in activation between 0-back and 3-back, and another image based on 0-back versus 2-back condition). These contrast images later will be entered into second-level random-effects analyses for between-groups (i.e., MA users versus control participants) and between-sessions (Phase 1 versus Phase 2) comparisons. In addition to straightforward univariate SPM analysis, which will elucidate the set of regions involved in working memory and selective attention, we will perform functional and effective connectivity analysis (looking at fronto-thalamo-striatal and fronto-limbic circuitry in particular based on our preliminary PET data). Functional connectivity is defined as the temporal correlations between spatially remote neurophysiological events (Friston et al., 1993a). There are now several methods available for elucidating patterns of correlated activity and determining their contributions to the observed functional connectivity, including principal components analysis (PCA) {Friston et al., 1994, REF#10408}, independent components analysis (ICA) {Keown and Sejnowski, 1998), and partial least squares (PLS; Mcintosh et al.,1996). Functional connectivity analyses describe correlations, but they do not provide insight about their origins. Path analysis, on the other hand can be used to test hypotheses about effective connectivity. Several path analyses of functional MRI (fMRI) data have reported (e.g., Büchel and Friston,1997; Bavelier et al., 2000; Bullmore et al., 2000). The PET image will be reconstructed using the ECAT v7.3 OSEM (3 iterations, 16 subsets) following the corrections applied for decay, attenuation, normalization, and scatter. Prior to further data processing the dynamic frames Page 16 of 37
  17. 17. Fallypride IND Application June 22, 2007 will be spatially aligned to the middle frame using SPM2 to correct for patient head motion. These dynamic datasets will then be used to create time–activity curves. The region of interests (ROIs) will be drawn on each subject’s T1 MRI and then the ROIs will be automatically transferred to the PET images using the transformation that coregisters the MRI to the PET images. The cerebellum (cerebellar lobes) is chosen as the reference region, having a minimal amount of specific binding to dopaminergic sites. The binding potential (BP) will be calculated from the distribution volume ratio (DVR) values using a graphical analysis (Logan Plot): BP = DVR-1. III. Chemistry, Preparation, and Control A. Chemistry and Manufacturing Introduction. Based on past experience and published data, the chemistry and manufacturing of both drug substance and drug product do not present signals of potential human risk. B. Drug Substance. B.1. A Description of the Drug Substance and Drug Product: The drug substance in the drug product [18F]Fallypride Injection is (S)-N-((1-Allyl-2- pyrrrolidinyl)methyl)-5-(3-[18F]fluoropropyl)-2,3-dimethoxybenzamide. This name will be abbreviated as [18F]Fallypride for this section of the IND application, and the chemical structure is shown in Figure 1. Figure 1: Structure of (S)-N-((1-Allyl-2-pyrrrolidinyl)methyl)-5-(3- [18F]fluoropropyl)-2,3-dimethoxybenzamide [18F]Fallypride H H O N N OCH3 CH CH CH2 2 CH2CH2CH2 OCH3 X X=19F X=18F Page 17 of 37
  18. 18. Fallypride IND Application June 22, 2007 Fallypride [18F]Fallypride C20H2919FN2O3 C20H2918FN2O3 Exact Mass: 364.22 Exact Mass: 363.22 Mol. Wt.: 364.45 Mol. Wt.: 363.46 m/e: 364.22 (100.0%), 365.22 (22.1%), 366.22 m/e: 363.22 (100.0%), 364.22 (22.5%), 365.23 (3.0%) (2.3%) C, 65.91; H, 8.02; F, 5.21; N, 7.69; O, 13.17 C, 66.09; H, 8.04; F, 4.95; N, 7.71; O, 13.21 For the manufacture of [18F]Fallypride Injection the terms "drug substance" and the "drug product" are essentially synonymous. Where appropriate, however, the drug substance will be discussed separately from the drug product. B.2. Characterization of the Drug Substance. B.2.a. Description Chemical Name: (S)-N-((1-Allyl-2-pyrrrolidinyl)methyl)-5-(3- [18F]fluoropropyl)-2,3-dimethoxybenzamide Chemical Formula: C20H2918F N2O3 Abbreviation: [18F]Fallypride The radionuclide, fluorine-18 (18F), decays by positron (β+) emission with a half-life of 109.8 min. The daughter product of the decay is the stable radionuclide, oxygen-18 (18O). B.2.b. Name and Address of the Drug Substance Manufacturer. Radiopharmaceutical Production / Cyclotron Facility Building 345 Veterans Administration Greater Los Angeles Health System 11301 Wilshire Blvd. Los Angeles, CA 90073 Page 18 of 37
  19. 19. Fallypride IND Application June 22, 2007 B.2.c. General Method of the Preparation of the Drug Substance. B.2.c.1. Production Radiochemical synthesis of [18F]Fallypride Injection 1, 2, 3 utilizes the Kryptofix® 222- assisted nucleophilic no-carrier-added [18F]fluorination of (S)-2,3-dimethoxy-5-[3-[[(4- methylphenyl)sulfonyl]oxy]propyl]-N-[[1-(2-propenyl)-2-pyrrolidinyl]methyl- benzamide, abbreviated as Tosyl-Fallypride (Mol Wt. 516.65, C27H36N2O6S), followed by HPLC purification (Figure 2). No-carrier-added [18F]fluoride is produced in the TR-19 cyclotron by proton bombardment of enriched [18O]water by the cyclotron facility. The manufacturing method, quality control, and information about stability of the drug substance will be discussed in the following section, Drug Product (Tables 2, 3, and 4). C. Drug Product. The individual dose of the drug product (Table 2) will be prepared in the form of a multi-dose batch (Table 3) as a sterile nonpyrogenic solution for intravenous injection. C.1. Composition of the Individual Dose of the Drug Product. Table 2: Composition of the Individual Dose of the Drug Product Component Quantity [18F]Fallypride ≤ 5 mCi Fallypride* ≤ 1.8 μg 0.9% NaCl < 12 mL * The amount of carrier [18F]Fallypride calculated, assuming a dose of 5 mCi and specific activity of 1000 mCi/μmol or greater, corresponds to a dose of <0.005 μmol/ 5 mCi; the maximum dose of carrier per 5 mCi = (0.005 μmol/dose) x (364.45 μg/μmol) = 1.8 μg/dose. 1 Mukherjee, J.; Yang, Z.Y.; Das, M. K.; Brown, T.; Fluorinated benzamide neutoleptics-III. Development of (S)-N-[(1-allyl-2-pyrrolidinyl)methyl]-5-(3-[18F]fluoropropyl)-2,3-dimethoxybenzamide as an Improved Dopamine D-2 Receptor Tracer. Nucl. Med. Biol. 1995, 22, 283-296. 2 Mukherjee, J.; Christian, B. T.; Dunigan, K. A.; Shi, B.; Narayanan, T. K.; Satter, M.; Mantil, J.; Brain Imaging of 18F-Fallypride in Normal Volunteers: Blood Analysis, Distribution, Test-retest Studies, and Preliminary Assessment of Sensitivity to Aging Effects on Dopamine D-2/D-3 Receptors Synapse, 2002, 46, 170-188. 3 Mukherjee, J.; Yang, Z.Y.; Brown, T.; Lew, R.; Wernick, M.; Ouyang, X.; Yasillo, N.; Chen, C. T.; Mintzer, R.; Cooper, M.; Preliminary Assessment of Extrastriatal Dopamine D-2 Receptor Binding in the Rodent and Nonhuman Primate Brains Using the High Affinity Radioligand, 18F-Fallypride. Nucl. Med. Biol. 1999, 26, 519-527. Page 19 of 37
  20. 20. Fallypride IND Application June 22, 2007 C.2. Batch Formula Quantity Table 3: Batch Formula Quantity Component Per Batch Per Dose Tosyl Fallypride 2.0-2.2 mg Removed during processing F-18 900-1100 mCi ≤5 mCi 0.9% NaCl 12 mL < 4 mL C.3. List of Components of the Drug Product by Name. 1) Kryptofix® 222, 4,7,13,16,21,24-Hexaoxa-1,10 diazabicyclo[8.8.8]hexacosane, F.W. 376.50 g/mol, Mp 70-73 °C. Acceptable Suppliers: Sigma-Aldrich, ABX 2) Acetonitrile, HPLC grade, F.W. 41.05 g/mol, Bp 81.6 °C, d20 0.786 g/ml. Acceptable Suppliers: Sigma-Aldrich, Mallinckrodt, Baker, Pierce. 3) Potassium Carbonate, 99.995%, F.W. 138.21 g/mol, Mp 891 °C. Acceptable Suppliers: Sigma-Aldrich 4) Water, HPLC grade, F.W. 18.02 g/mol. Acceptable Suppliers: VA Radiopharmaceutical lab, produced with Millipore MilliQ system. 5) Helium Gas, He, Research Grade or Ultra-High Purity. Acceptable Suppliers: Matheson, Air Gas, Praxair. 6) (S)-2,3-dimethoxy-5-[3-[[(4-methylphenyl)sulfonyl]oxy]-propyl]-N-[[1-(2- propenyl)-2-pyrrolidinyl]methyl-benzamide, abbreviated as Tosyl-Fallypride, C27H36N2O6S, F.W. 516.65 g/mol, Identity by 1H -NMR. Acceptable Suppliers: ABX. 7) Anion-exchange cartridge (for 18F- trap). Acceptable Suppliers: ABX. 8) µBONDAPAK-CN RP, 10µ, HPLC semi-prep column (7.8 x 300 mm). Acceptable Suppliers: Waters. 9) Spherisorb-C18 S5ODS2, 5µ, HPLC analytical column (4.6 X 250 mm) with 3 cm guard column. Acceptable Suppliers: Waters. 10) Membrane Sterilizing Filter, 0.22 μm pore size, vented, sterile. Acceptable Suppliers: Gelman, Millipore (Millex-GS 0.22 µm). 11) Normal Saline, 0.9%, USP; clear colorless solution packaged in sterile rubber- stoppered vials. Sterile and pyrogen free. Acceptable Suppliers: Abbott, Fujisawa, Kendall-McGaw. 12) Vial, empty, sterile, nonpyrogenic, 20 or 30 mL. Acceptable Suppliers: Abbott Laboratories, Lyphomed, Wheaton, EP Scientific. 13) [18F]Fluoride ion/[18O]Water, [18F]F-/[18O]H2O (cyclotron target product). Acceptable Suppliers: VA radiopharmaceutical lab. Page 20 of 37
  21. 21. Fallypride IND Application June 22, 2007 C.4. Preliminary Production Data. The theoretical decay corrected yield is 100%. Actual yields in radiochemical synthesis are lower and can be expected in the range of 5 – 25% (decay corrected). The overview of the manufacturing results and Quality Control (QC) procedures are presented in Table 4. Page 21 of 37
  22. 22. Table 4: [18F]Fallypride Synthesis Process Results [F-18] Yield (% - specific Kryptofix Radichemical LAL pyrogen Date Fallypride non decay activity pH sterility Purity (%) test (EU/mL) (mCi) corrected)) (Ci/umol) (µg/mL) 7/5/06 26 2.94 3.81 99.60 <50 µg/mL 5.73 <0.125 EU/mL No Growth 7/19/06 57 5.50 2.01 100.00 <50 µg/mL 5.90 <0.125 EU/mL No Growth 8/9/06 10 1.04 -- 100.00 <50 µg/mL 4.50 <0.125 EU/mL No Growth 8/16/06 202 18.57 2.82 100.00 <50 µg/mL 5.00 <0.125 EU/mL No Growth 8/22/06 183 18.96 20.97 100.00 <50 µg/mL 5.00 <0.125 EU/mL No Growth 8/23/06 213 19.91 -- 100.00 <50 µg/mL 5.00 <0.125 EU/mL No Growth 9/6/06 226 23.06 -- 100.00 <50 µg/mL 5.00 <0.125 EU/mL -- 9/20/06 162 16.22 4.05 100.00 <50 µg/mL 5.00 <0.125 EU/mL No Growth 9/26/06 154 16.68 2.66 100.00 <50 µg/mL 5.30 <0.125 EU/mL No Growth 9/27/06 203 16.28 14.63 100.00 <50 µg/mL 5.00 <0.125 EU/mL No Growth 10/10/06 164 16.96 9.84 100.00 <50 µg/mL 5.00 <0.125 EU/mL No Growth 10/11/06 134 12.28 4.61 100.00 <50 µg/mL 4.70 <0.125 EU/mL No Growth 10/31/06 236 19.62 32.22 100.00 <50 µg/mL 4.90 <0.125 EU/mL No Growth 11/1/06 200 18.62 12.39 99.80 <50 µg/mL 4.50 <0.125 EU/mL No Growth 11/7/06 196 18.01 6.35 100.00 <50 µg/mL 4.70 <0.125 EU/mL No Growth 11/14/06 192 18.03 17.30 99.80 <50 µg/mL 4.70 <0.125 EU/mL No Growth 11/15/06 248 23.13 13.60 100.00 <50 µg/mL 4.80 <0.125 EU/mL No Growth 12/12/06 273 25.42 16.42 100.00 <50 µg/mL 5.00 <0.125 EU/mL No Growth 1/23/07 287 26.95 11.49 100.00 <50 µg/mL 5.60 <0.125 EU/mL No Growth 2/21/07 303 28.05 20.71 100.00 <50 µg/mL 5.50 <0.125 EU/mL No Growth
  23. 23. 3/20/07 240 24.32 7.92 100.00 <50 µg/mL 5.50 <0.125 EU/mL No Growth 3/28/07 354 29.65 22.38 100.00 <50 µg/mL 5.30 <0.125 EU/mL No Growth 4/3/07 210 19.85 5.46 100.00 <50 µg/mL 5.40 <0.125 EU/mL No Growth 4/4/07 83 8.30 8.89 98.96 <50 µg/mL 5.50 <0.125 EU/mL No Growth 4/11/07 134 13.33 5.07 99.25 <50 µg/mL 5.00 <0.125 EU/mL No Growth 4/18/07 261 25.51 15.16 99+ <50 µg/mL 5.00 <0.125 EU/mL No Growth 5/17/07 135 12.65 3.63 100.00 <50 µg/mL 5.00 <0.125 EU/mL No Growth 6/6/07 60 6.47 4.20 99+ <50 µg/mL 5.30 <0.125 EU/mL No Growth The procedure described below was applied in 30 preparations of [18F]Fallypride for tests and in vitro studies. The average yield of the labeled product was 11.6% of the starting material (900-1100 mCi). The drug product met specific activity specification requirements of this IND (greater than 1000 mCi/μmol). Radiochemical purity was greater than 98% and pH was in the range of 4.0 – 8.0 in all cases. Content of the catalyst, Kryptofix® 222, remaining in the drug product was below the detectability level (< 50 μg/mL or < 1 mg/batch). All randomly analyzed lots passed the sterility tests (Fluid Thioglycollate and Soybean-Casein Digest) and the Endotoxin test (<0.125 EU/mL or <2.5 EU/batch). C.5. Facilities for Preparation and Quality Control. C.5.a. Facilities. Drug product is manufactured and packaged at the Radiopharmaceutical Production / Cyclotron Facility of West Los Angeles VA Medical Center, Building 345. Access to work areas, production and testing equipment, components, containers and closures, and the PET drug products, is restricted to authorized personnel. C.5.b. Approved Components Area. Approved components are stored in a designated area segregated from the general stock and are distinctively labeled. C.5.c. Critical Operation Area and Critical Handling. Aseptic operations with sterile components are carried out in a laminar hood (rating of Class 100). The laboratory is kept free of extraneous materials. Traffic in the area is restricted to authorized personnel.
  24. 24. C.6. Manufacturing Steps. Cyclotron production of [18F]fluoride in the Advance Cyclotron Systems TR19. The nuclear process responsible for the production of the [18F]fluoride ion is the 18O(p,n)18F reaction. The target material is the stable isotope, oxygen-18, which is in the form of [18O]water. Production of [18F]fluoride ion is achieved by the bombardment of the [18O]water target material with 17.5 MeV protons. Proton beam parameters are: Beam Current = 35 µA; time of bombardment = 20 – 30 min. The target yield is variable and can be in range of 900 – 1200 mCi. The volume of the target material used during a bombardment is 1.7 mL. The target body is made of Niobium, and the target window is made of a thin Havar foil. The target body is cooled with chilled water, and the target window is cooled with a stream of Helium gas. Step #1. The recovery of [18F]fluoride. The recovery of [18F]fluoride is performed by passing the radioactive target material (900 - 1200 mCi [18F]fluoride in 1700 μL [18O]water) through a anion-exchange cartridge followed by extraction of the radioactivity with a solution of potassium carbonate (3.0 mg) in water (0.3 ml), and transfer from the anion-exchange to a cylindrical Pyrex glass vessel #14, which is a part of the radiochemistry set-up. Step #2. Preparation of anhydrous complex K/K222)+/[18F]F-. K2.2.2 (10 mg) is dissolved in 1 mL of CH3CN. Kryptofix solution is added to the vessel containing the activity. The solution is heated and water is removed by azeotropic drying. Azeotropic evaporation of the residual water is continued by addition of two more 1.0 mL portions of CH3CN. Upon completion of the stage, [18F]fluoride ion exists in the form of the anhydrous complex [K+ · Kryptofix® 222] · [18F]F–. Step #3. Radiofluorination. A solution of Tosyl Fallypride (2.0 – 2.2 mg) in anhydrous CH3CN (2 mL) is added to Vessel and heated in a microwave cavity for 1.5 minutes. After a short cooling period, volatiles are evaporated. Step #4. Purification of the [18F]fluorinated products. The residue is taken up in ethanol and the solution is passed through a series of Sep- Pak purification cartridges (IC-H Light, QMA Plus, and Alumina-N Plus) to a second reaction vessel. The solution is evaporated to near dryness, diluted with 2.0 mL of preparative HPLC mobile phase (HPLC solvent for the preparative column [70% 10mM NaH2PO4 (pH = 3.50) and 30% ethanol]), injected onto the HPLC column, and eluted at a flow rate of 3 mL/min. 4 All production glassware is nonpyrogenic..
  25. 25. The radioactive peak of [18F]Fallypride, with a retention time of RT ≈ 13min corresponding to the “cold” Fallypride standard, is collected into an empty flask. Step #5. Formulation of the final product. The solution in the rotary-evaporator flask is diluted with 0.9% saline solution and filtered through a sterile filter unit into a sterile vial yielding the [18F]Fallypride Injection. Step #6. Sampling for QC. The vial containing the Drug product is transferred into the laminar fume hood (class 100). A QC sample of the drug product (about 0.8 mL) is taken aseptically by the sample syringe. C.7. Quality Control. C.7.a. Proposed Acceptable Limits and Analytical Methods. 1) Color and clarity: acceptable limits: colorless, clear liquid method: visual observation 2) pH: acceptable limits: 4 – 8 method: application of drug product solution to pH paper strips 3) Total multi dose batch radioactivity assay: acceptable limit: > 10 mCi method: assay in a Capintec dose calibrator instrument: Capintec dose calibrator (CRC-35R). The dose calibrator Capintec CRC-35R is NIST-calibrated. Due to the short half life of 18F, NIST traceable sources of 18F are not available. For this reason, the accuracy of the dose calibrator is verified annually with two NIST-traceable sources that became conventional for PET, 137 Cs (662 keV) and 57Co (with gamma energy lower than 511 keV). 4) Specific activity: acceptable limit: > 1000 mCi/µmol method: An aliquot of the drug product (100 µL) of known radioactivity is applied to HPLC column Spherisorb-C18 5µ ODS2; Mobile Phase: 10 mM NaH2PO4 (pH
  26. 26. 3.5) and CH3CN (35: 65 v/v) at a flow rate of 2 mL/min is used to elute the radioligand, which has a retention time of 3.7 – 4.8 min. The area of the UV absorbance peak measured at 254 nm corresponding to carrier product is measured and compared to a standard curve relating mass to UV absorbance. instruments: Capintec dose calibrator (CRC-35R), Waters chromatograph with a UV detector (254 nm), and a Radiation detector. 5) Radiochemical purity acceptable limit: > 95% method: assay by HPLC; Spherisorb-C18 ODS2 (5m; 4.6 x 250 mm; Waters). 70% 10mM NaH2PO4 (pH = 3.50) and 65% acetonitrile 6) Chemical purity (Kryptofix® 222): acceptable limit: < 50 µg/mL or 1 mg/batch method: assay by TLC 7) Endotoxin limits acceptable limit: 175 Endotoxin Units per patient dose. Because an entire batch can be used as a single dose, the Endotoxin limit is 175 Endotoxin Units per batch. method: LAL test 8) Sterility testing acceptable limits: meets criteria under Sterility Tests in USP (Sterility tests, USP 28). method: Fluid Thioglycollate and Soybean-Casein Digest Sterility tests are performed by the microbiology Laboratory of West Los Angeles VA Medical Center. C.7.b. Quality Control Schedule. Due to the short half-life of 18F (109.8 min), the quality assurance of dedicated batches of the Drug Product will be performed based on the following schedule: 1) Color and clarity – every production batch before the product vial is released for injection 2) pH – every production batch before the product vial is released for injection
  27. 27. 3) Total radioactivity assay – every production batch before the product vial is released for injection 4) Specific activity – every production batch before the product vial is released for injection 5) Radiochemical purity – every production batch before the product vial is released for injection 6) Chemical purity (Kryptofix® 222) – every production batch, ASAP after the product vial is released for injection 7) Bacterial endotoxin (LAL test) – every production batch, ASAP after the product vial is released for injection. We stipulate such a schedule due to the short half-life of 109.8 min. The short half-life causes natural radioactive decay of the Drug Substance and a decline of the specific radioactivity during the QC test. The LAL test takes 1 h. 8) Sterility test – every production batch, ASAP after releasing product vial for injection. C.7.c. Information to Support the Radiochemical Stability of the Drug Product. The Radio HPLC analysis of the Drug Product at 0.1, 2 and 6 h after the end-of-the-synthesis (EOS) did not show any decomposition of the Drug Substance. The radiochemical purity at all time points was 98 – 99%. C.7.d. Expiration Time. Expiration time will be determined based on the product radiochemical stability and the specific activity. The expiration time will not exceed 6 h because the product stability has been tested only for 6 h. Due to radioactive decay, the specific activity of the drug product decreases during storage time. At time of injection, the specific activity of the drug product must be greater than 1000 mCi/µmol. Expiration time of a multi-dose batch will be determined individually for each batch based on the specific activity expiration time and radiochemical stability expiration time, whichever comes first. C.7.e. Labels.
  28. 28. [F-18]Fallypride Lot No.:____________ Sterile, nonpyrogenic solution for I.V. administration. New drug limited by Federal law to investigational use only VA GLAHS Cyclotron Facility/Radiopharmaceutical Laboratory Activity ________mCi; Volume:__________mL; Time___________ Specific Activity_________________mCi/mmol; Time___________ Expires:_________ h after specific activity determination The final product container bears the label shown above. C.7.f. Manufacturing Responsibility. By adhering to the procedures given in this section, we will maintain appropriate laboratory standards of identity, purity, and quantitative composition of the Drug Product to allow safe, reliable PET research. The supervision of the Drug Product manufacture is the responsibility of Judah Farahi, Ph.D. IV. Environmental Assessment. A categorical exclusion is requested regarding the environmental assessment. Activity excreted by the subjects is disposed through the sewage system at such low amounts that there will be no detrimental effects to the environment. All potentially contaminated laboratory materials are held at least 10 half-lives for decay. In addition, these materials are surveyed to ensure background levels of activity prior to disposal through regular waste systems. V. Pharmacokinetics and Biodistribution Studies in Animals. In addition to studies reported in Dr. Kessler’s IND, Jogeshwar Mukherjee, PhD has reported several studies on the use of [18F]fallypride in nonhuman primates. 1. Quantitation. Dr. Mukherjee has studied the kinetics of brain uptake and used several methods to quantify D2 receptors in brain (Christian et al 2000). An arterial input function was used to calculate receptor levels in striatal and extrastriatal regions with compartmental analyses and Logan plots. All methods
  29. 29. were in close agreement, including the use of the cerebellum as a reference tissue instead of the arterial input function. 2. Occupancy by Antipsychotic Medications. The occupancy of D2 receptors in brain was examined in both rats and nonhuman primates (Mukherjee et al 2001). Some prior studies with SPECT D2 receptor imaging reported that atypical medications like clozapine had preferentially greater occupancy of extrastriatal than striatal receptors (Pilowsky et al 1997). In contrast, using carefully controlled conditions in monkeys, Mukherjee et al. (2001) found equal occupancy of striatal and extrastriatal D2 receptors by typical and atypical antipsychotic medications. 3. Displacement by Endogenous Dopamine. Because fallypride has high affinity for the D2 receptor (Kd ~ 30 pM), many scientists would predict that it could not be displaced by amphetamine-induced dopamine release. In fact, amphetamine (0.75 – 1.5 mg/kg I.V.) caused significant displacement of [18F]fallypride binding in rhesus striatum (Mukherjee et al 1997). Marc Laruelle, MD (Columbia University) and Robert Kessler, MD (verbal reports at meetings) have had similar results. Thus, affinity is not the only determinant of displacement by endogenous neurotransmitter. Two other factors that may be more important are the off-rate of binding and the lipophilicity of the radiotracer. VI. Pharmacology and Toxicology of [18F]-Fallypride Data Supplied by Dr. Robert Kessler Fallypride is a selective, high affinity D2 receptor antagonist with KD of 30 pM (Christian et al, 2000; Kessler et al, 1993). Pharmacological doses of this agent would be expected to show typical D2 receptor antagonist activities of extrapyramidal side effects, sedation, and catalepsy. However, the proposed dose is well below that of pharmacological effects. Thus, the current study is not expected to cause any effects or side effects. Robert Kessler, MD has an active IND for [18F]fallypride (also called N-allyl-5- fluoropropylepidepride). Dr. Kessler has authorized the FDA to review his IND on behalf of the current proposal for all information regarding pharmacology of this tracer, including toxicity, pharmacokinetics, safety, and efficacy. Please note that Dr. Kessler’s IND was approved for the use of a mass dose of no greater than 0.9 µg, calculated as follows: 5 mCi * 1 µmol/2,000 mCi * 364 µg/µmol = 0.9 µg Since the time of original submission, Dr. Kessler has studied many human subjects and found no pharmacological effects or side effects. In a study carried out by our lab at UCLA a maximal dose of 2 µg, the dose currently being requested, corresponding to about 0.03 µg/kg administered to a 70 kg adult, has also produced no pharmacological effects or side effects. With this information, we request a maximal mass dose of 2 µg per scan (2 scans maximum) per participant. We have included Dr. Kessler’s original IND submission has information on Pharmacology and Toxicology (Appendix B, continued.). His data support a 100-1,000-fold safety factor for the proposed dose (0.04 µg/kg) relative to the minimum effective dose in animals. The Pharmacology and Toxicology Data in Dr. Kessler’s IND application can be summarized as follows:
  30. 30. 1) Acute Toxicity in Rats. The maximal dose in rats (600 µg/kg) caused mild sedation, no deaths, and no abnormalities on postmortem examination. This dose is approximately 20,000 fold higher than the proposed human dose by us at UCLA (0.03 µg/kg). 2) Subacute Toxicity in Rats. Ten consecutive daily doses of fallypride (1.5 and 6 µg/kg I.V.) showed no pathological effects on postmortem examination. The higher dose (6 µg/kg) is ~200 fold higher than the proposed human dose. 3) Subacute Toxicity in Rhesus Monkeys. Ten consecutive daily doses of fallypride (3 µg/kg I.V.) showed no significant effect on blood chemistries, CBC, and chest X-ray. This dose (3 µg/kg) is 100 fold higher than the proposed human dose. 4) Pharmacological Effects. The lowest dose in monkeys to produce the earliest observable pharmacological effects was 2.2 nmol/kg, which corresponds to ~0.8 µg/kg I.V. The behavioral effect was decreased pacing and decreased reactivity to eye contact. The HED (Human Equivalent Dose) would be 0.26 µg/kg (= 0.8 X 0.324). The maximal proposed human dose is 0.029 µg/kg (= 2 µg / 70 kg subject). Thus, the proposed human dose is 9 times lower than the minimally effective behavioral dose in monkeys. For this reason and the fact that the proposed dose is only two fold higher than that used by Dr. Kessler in humans with no effects, we expect that our studies will show no pharmacological effects. Dr. Kessler also calculated that a fallypride dose of ~0.015 µg/kg would occupy ~0.7% of striatal D2 receptors. Since this low dose is in the linear range of receptor occupancy, our proposed dose (0.03 µg/kg) would occupy 1.4% of receptors, at least 30 fold lower than any side effects. 5) Radiation Dosimetry. Rat biodistribution data were used to calculate organ dosimetry. A dose of 5 mCi was estimated to cause the following exposures (rem): 0.8 to red marrow; 0.25 to spleen; 0.1 to testes, 0.19 to whole body, and 1.05 to small intestine. These exposures are within guidelines for research studies. After the original IND application, Dr. Kessler submitted additional data on dosimetry based upon human whole body imaging. The tabulation of radiation doses to participants is included. Studies are also available for the lethal dose in rats (LD50) of unlabelled fallypride as reported by Dr. J. Mukherjee (University of California – Irvine). LD50 has been evaluated to be 225 µmol/kg, which is 90 mg/kg of fallypride (molecular weight of fallypride hydrochloride is 400 mg/mmol). If this rat toxicity were to be extrapolated to humans, then a total dose of unlabeled fallypride lethal in humans (for an average weight of 70 kg) would be 90 mg X 70 kg = 6.3 g. The maximal amount of fallypride that we propose to be administered in our PET studies is 2 µg. Since the LD50 dose of unlabeled fallypride in humans (for an average weight of 70 kg) is approx 6.3g, the total dose of fallypride proposed for our study is approximately 2 X 106 times below its acute toxicity levels. As previously mentioned in the section regarding the tests which will be used to establish the identity, purity, and non-toxicity of the drug (Question 6b) we will also perform a chromatography analysis to assess the specific activity of the radiolabeled benzamide after every radiosynthesis. Based on both the reported results in the literature and our laboratory’s 18F radiolabeling results, we have set a lower limit of 1,000 mCi / 1 µmol for the specific activity of 18F-fallypride. Expressing in µg, 1,000 mCi / 364 µg giving 5 mCi / 1.82 µg as the maximal activity (mCi) proposed to be injected into any human participant. VII. Dosimetry.
  31. 31. Tabulation of Radiation Doses to Subjects Radioactive Material or Procedure: [18F]fallypride Administered Quantity or View of Exposure: 5 mCi Subject Age: 18 – 45 (four PET studies) ORGAN RADIATION DOSE (rem) Per single Administration [Per Year (12 mos.)] Adrenals 0.27 [1.1] WHAT IS GOING ON WITH THE 2 NUMBERS?, IS THE NUMBER ON THE LEFT “PER SINGE ADMINISTRATION & THE ONE ON THE RIGHT PER YEAR? IF SO, HOW ABOUT SETTING OFF THE “PER YEAR DESIGNATION WITH SQUARE BRACKETS – IN THE COLUMN HEADINGA ND ALSO IN THE CELLS OF THE TABLE. Brain 0.165 [0.66] Breasts 0.13 [0.52] Esophagus 1 0.165 [0.165] Gallbladder Wall 2.2 [8.8] GI-tract: Lower Large Intestine 0.18 [0.72] Small Intestine 0.19 [0.76] Stomach 0.19 [0.76] Upper Large Intestine 0.205 [0.82] Colon2 0.19 [0.78] Heart Wall 0.9 [3.6] Kidneys 0.22 [0.88] Liver 1.3 [5.2] Lungs 0.55 [2.2] Muscle 0.145 [0.58] Ovaries 0.185 [0.74] Pancreas Red Marrow 0.36 [1.4] Bone Surfaces 0.28 [1.12] Organ RADIATION DOSE (rem) Per single Administration Per Year (12 mos.) Skin 0.1 0.4 Spleen 0.75 [3]
  32. 32. Testes 0.13 [0.52] Thymus 0.165 [0.66] Thyroid 0.12 [0.48] Urinary Bladder Wall 3 1.65 [6.6] Uterus 0.235 [0.94] Lens of Eyes4 0.1 [0.4] EFFECTIVE DOSE 0.39 [1.56] NOTES: 1 Since no dose is explicitly tabulated for esophagus, thymus dose is used (as per ICRP 80) 2 Colon Dose estimated by [0.57 (DoseULI) + 0.43 (DoseLLI)] (as per ICRP 80). 3 Dynamic urinary bladder model used; void interval of 2 hours 4 Assumed to be the same as the dose to the skin DOSIMETRY SOURCE: Attached communication from RM Kessler, MD. NOTES: A. If more than one agent or radiation source is to be used, fill out additional copies of this tabulation (see attachment B). B. If radiographic studies are involved, indicate primary site(s) or organs to be irradiated and include skin or "entrance" dose. C. Estimates of doses for the organs and tissues listed in the table should be as accurate as possible. If this is inappropriate for some sites, e.g., for those that are distant from the irradiated site in radiographic studies, note the basis for dose estimates in the protocol or leave blank, as appropriate. IX. Human Experience. Dr. Kessler has injected both healthy subjects and schizophrenic patients with [18F]fallypride. As of December 6, 2002, Dr. Kessler had injected [18F]fallypride in 63 human subjects and never found any subjective reports of effects, nor any clinically significant changes in vital signs or blood chemistries. See Dr. Kessler's letter in Appendix E. As part of in Phase I IND study, Dr. Kessler collected human biodistribution data to calculate organ dosimetry. The results are detailed in Appendix G "Report of Phase I IND for [18F]N-allyl-5- fluoropropylepidepride." The three organs with highest radiation burden (rem/mCi) were: gallbladder wall: 0.44; lower large intestine wall: 0.36; and urinary bladder wall: 0.33. As a measure of overall exposure, the effective dose was 0.078 rem/mCi. Dr. Mukherjee has recently reported the results of a detailed test/retest kinetic analysis of [18F]fallypride brain uptake and metabolism in six healthy subjects (Mukherjee et al 2002). His study demonstrates the validity of using a reference tissue model to assess receptor binding in brain. We plan to use such a reference tissue method in the current study, which will obviate the need for arterial blood sampling and radiometabolite analysis.
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