Clinical trial NCT04015869
Conditions: Alcohol Use Disorder
Sponsor: Yale University
Start date: 2019-07-01; Completion date: 2023-09-01
Enrollment: 10 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
The goal of this study is to determine whether intravenous infusion of allopregnanolone (ALLO) attenuates stress-induced craving and stress-induced anxiety in a clinical laboratory setting. The secondary objective of this project is to characterize the behavioral effects of ALLO in heavy drinkers.

Detailed description:
This is a double-blind, placebo-controlled study that will randomize heavy "at risk" drinkers to 2 groups: targeted dose of 100 nM of ALLO or placebo. Participants will receive a continuous infusion of ALLO (or placebo) for approximately 175 min. Sixty minutes after the start of the infusion, when ALLO levels are relatively stable, the stress induction paradigm will be introduced. Participants will be presented with a stress cue and a neutral cue in random order. Stress and neutral cues will consist of personalized 5-minute scripts created prior to testing. Measures of stress-induced craving and stress-induced anxiety will be presented before the cue (pre), immediately following the cue (post) and 10 minutes after (recovery). An alcohol infusion (40mg%) begins at approximately 120 minutes into ALLO Infusion. Once the alcohol infusion has reached the targeted dose of 40mg% the infusion is clamped for approximately 20 minutes. The study outcomes will include measures of stress-induced craving and stress-induced anxiety as well as measures of subjective mood effects, cognitive performance, and motor coordination.

Arms: arm 1: Allopregnanolone infusion dose of 100nM over 175 min arm 2: Placebo infusion
Interventions: intervention 1: Allopregnanolone intervention 2: Placebos
Intervention descriptions: intervention 1: neurosteroid intervention 2: placebo

Protocol documents:
Prot_SAP_001.pdf: 
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PROJECT DESCRIPTION 
 
Title: Effect of allopregnanolone on stress-induced craving  
 
1.  Principal Investigator:  
Elizabeth Ralevski, PhD 
 
Co-Investigator: Ismene Petrakis, MD 
 
1A. Authorized Prescribers: Ismene Petrakis, MD; Gihyun Yoon, MD 
 
2.  Purpose:  
 
The role of stress in the vulnerability, initiation, and maintenance of alcohol use disorders (AUDs) is 
supported by a rich literature and represents a clinically important area of study. The dysregulation of 
the stress response is seen as critical in the development and maintenance of AUD. Neurosteroids are 
considered central in the regulation of the stress response and the hypothalamic pituitary adrenal 
(HPA) axis, and have robust direct central nervous system (CNS) effects, as well as anxiolytic and 
neuroprotective effects. Neurosteroid modulation of the stress system has been identified as an 
important key to understanding the mechanisms of how stress-system dysregulation may lead to the 
development of AUD.    
  
Neurosteroids act on a number of different receptors; most notably those that regulate CNS activity, 
gamma-amynobutyric acid (GABA) and glutamate, but also nicotinic receptors, glycine receptors and 
calcium and potassium channel receptors. However, their potent actions on GABAa receptors is of 
particular importance because it is through these most widely disseminated receptors in the brain 
that neurosteroids are believed to exert their powerful anxiolytic, analgesic, antiepileptic and 
neuroprotective properties. Of all neurosteroids, alloprognanolone (ALLO) is the most direct 
modulator of GABAa receptors and it is this action that has sparked the interest in ALLO’s relevance 
for a number of stress-related psychiatric conditions. Considerable evidence suggests that ALLO 
mediates alcohol’s subjective stimulant and sedative effects, tolerance, dependence and withdrawal. 
The effects of ALLO administration have not been studied in humans in relationship to 
drinking outcomes and ALLO has not been administered to individuals with AUD.  
 
The goal of this Phase I study is to determine whether intravenous infusion of ALLO 
attenuates (1) stress-induced alcohol craving, stress-induced anxiety and (2) 
subjective stimulant/sedative effects of alcohol in individuals with AUD using a 
laboratory paradigm. The secondary objective of this project is to characterize the behavioral 
effects of ALLO in AUD. We propose a double-blind, randomized, between subject, placebo-controlled 
study in 60 individuals with AUD to compare one dose (1.5 mg/ml solution) of continuous (200 min.) 
infusion of ALLO (targeted plasma of 100 nM [IV bolus of 191 mcg/kg over 60 min. followed by 
continuous infusion of 57.4 mcg/kg/hr x 140 min.] n=30) to placebo (n=30). On a single test day, 
after 60 min of infusion - when ALLO levels stabilize - stress and neutral cues consisting of 
personalized 5 min. scripts will be presented in random order. The use of personalized scripts has 
been shown to produce robust and reliable craving and anxiety in subjects including heavy 
drinkers[1], alcohol dependent individuals (see the preliminary data section)[2] both with and 
without psychiatric comorbidity (including work by our group)[3-5] and, in those with other 
substance use disorders [6]. All participants will also receive alcohol administered intravenously 
using a clamp procedure, targeting a breath alcohol concentration (BrAc) of 40mg% (40 mg/dL).  
Alcohol will be administered following script presentation (20 min to target and clamped for 


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additional 30min). The main outcomes are measures of stress-induced alcohol craving, stress-
induced anxiety, and subjective alcohol effects; other secondary outcomes include subjective mood 
effects, cognitive performance, and motor coordination. 
 
Hypothesis 1 (Specific Aim 1): To determine if ALLO (targeted plasma of 100 nM), compared to 
placebo: 
Aim #1a: attenuates stress-induced craving for alcohol, measured by the Alcohol Urge Questionnaire 
(AUQ).   
Aim #1b: attenuates stress-induced anxiety, measured by the State Trait Anxiety Inventory (STAI-6).   
Aim #1c: attenuates the subjective stimulant/sedative alcohol effects (target BrAC=40 mg%), 
measured by the Biphasic Alcohol Effects Scale (BAES).  
 
Hypothesis 2 (Specific Aim 2): To characterize ALLO’s behavioral effects: 
Aim #2: to describe ALLO’s subjective mood effects, we will use Differential Emotions Scale (DES-
R), the Addiction Research Center Inventory (ARCI) and the Similarity to Drugs of Abuse Scale 
(SDAS). 
To evaluate the effects of ALLO administration on cognition and motor coordination: 
Aim #3a: to describe ALLO’s effects on cognitive functioning, we will assess verbal memory using 
the Hopkins Verbal Learning Test-Revised (HVLT-R), and response inhibition using the Go No-Go 
task.  
Aim #3b: to describe ALLO’s effects on motor coordination, we will use the Grooved Pegboard Test. 
 
Exploratory Aims: 
Aim #4: For all the above aims, we will also examine if the effects of ALLO are gender specific. 
 
 
3-4. Background and Significance  
SIGNIFICANCE: Alcohol use disorders (AUDs) have been characterized as stress-related conditions 
because the dysregulation of the stress response is seen as critical in their development and 
maintenance. The stress response is a physiologic multi-level and multi-system cascade of events that 
arises each time a stressful event occurs. However, chronic stress and repeated challenges to the 
stress system result in the dysregulation of this highly integrated network. In the last few decades, a 
growing body of evidence suggests the importance of neurosteroid modulation of the stress system, 
and this has been identified as an important key to understanding the mechanisms of how stress-
system dysregulation may lead to the development of AUD.   
 
Neurosteroids: The hypothalamic pituitary adrenal (HPA) response to stress, and the dysregulation 
of this response in psychiatric disorders has been well characterized[7-10].  More recently, a growing 
literature has studied the regulation of the HPA axis response by stress-derived neurohormones and 
their neuroactive metabolites, also known as neurosteroids [11]. These are derived from cholesterol, 
are synthesized both centrally and peripherally, have direct action in the brain and spinal cord, are 
potent modulators of neurotransmitters including gamma-amynobutyric acid (GABA) and glutamate, 
and are involved in cognition, arousal, motivation and emotion[7]. While they act on a number of 
different receptors, most notably GABA and glutamate, they are also believed to act on nicotinic 
receptors, glycine receptors and calcium and potassium channel receptors. Nevertheless, their potent 
actions on GABAa receptors seem to be of particular clinical relevance and are thought to be key to 
their anxiolytic, analgesic, antiepileptic and neuroprotective properties. Neurosteroids include 
pregnanolone and alloprognanolone (ALLO), and their actions are of interest in the regulation of 
stress disorders and are relevant for a number of psychiatric disorders including anxiety, addictive, 
and mood disorders, and disorders of cognition including traumatic brain injury (TBI)[7-10, 12]. 


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Compelling evidence indicates that neurosteroids, and ALLO in particular, have a significant 
therapeutic potential in psychiatric disorders. 
 
Neurosteroids in alcohol drinking behavior and AUD: A substantial body of preclinical and 
human studies show that neuroactive steroids have a role in alcohol reinforcement, tolerance, 
dependence and withdrawal. Data also indicates that neurosteroids, and ALLO in particular, may 
have significant therapeutic potential in AUD because of their ability to normalize a dysregulated HPA 
axis response typically found in chronic alcohol abuse. A2a. In preclinical studies: Alcohol increases 
the levels of progesterone (PROG) and ALLO in alcohol-preferring and non-preferring rats [13-15]. 
Administration of ALLO increases alcohol self-administration in non-dependent rats and in non-
dependent alcohol-preferring rats. However, chronic ethanol consumption in rats leads to decreases 
in plasma neurosteroid release [16-18], and decreased levels of ALLO after reintroduction of alcohol 
in alcohol-withdrawn animals[19]. Similarly, female monkeys who drink excessively tend to 
experience disruptions in their menstrual cycle, putatively due to neurosteroid depletion and low 
levels of ALLO [20]. Rodents who consume high doses of alcohol for long periods of time show 
decreases in alcohol consumption after a single infusion of ALLO [19, 21] suggesting ALLO may play 
an important compensatory role in mediating alcohol effects. A2b. In healthy humans: Similar to 
preclinical studies in healthy humans: a) alcohol intoxication increases plasma levels of ALLO [22, 
23], b) ALLO levels are correlated with levels of intoxication [22, 23] and subjective effects (“liking” or 
“wanting” more alcohol)[24], c) pretreatment with finasteride (reduces the formation of ALLO and 
PROG) blocks subjective stimulant effects of alcohol[24] and reduces drinking in men[25], and a 
single dose of dutasteride attenuates subjective sedative effects in non AUD men [26], d) women are 
less sensitive to the effects of alcohol and drink more during the follicular phase of their cycle, when 
ALLO levels are at their lowest [27]. There are some negative studies that show no relationship 
between intoxication and ALLO levels at lower doses of alcohol [28], in women in the follicular 
phase[29] or the luteal phase [30], and find lower levels of PROG and ALLO after alcohol 
administration [31]. A2c: In humans with AUD: Mirroring preclinical studies, women who drink 
excessively tend to experience disruptions in their menstrual cycle presumably due to neurosteroid 
depletion and lower levels of ALLO[32]. Dysregulation in HPA axis function has been reported in 
individuals with AUD while they are actively drinking, during withdrawal, and during abstinence [33-
35]. Also, suppression of neurosteroid release has been reported in individuals with AUD as well as 
their relatives after administration of naloxone [35, 36]. A recently published study examined the 
effect of progesterone stimulated ALLO in cocaine dependent men and women on stress-induced 
craving, mood, and cognitive functioning. High levels of ALLO were associated with decreased 
craving, increased mood ratings, and improved cognitive performance suggesting ALLO may have 
important therapeutic utility in addiction. In conclusion, data from animals and humans show that 
ALLO may have a significant therapeutic potential although there are no studies to date that 
examine the role of ALLO administration in AUD.   
 
ALLO administration in humans: ALLO has been available in Europe for years and has been 
safely administered to healthy men and women and to women with premenstrual dysphoric disorder 
(PMDD) [37-40]. In healthy women, a total cumulative dose of 0.09 mg/kg increases sedation and 
decreases saccadic eye movement [40]. The same dose administered to both men and women 
(adjusted for weight) produces more sedation in men, although the saccadic eye velocity was 
significantly more decreased in women [39]. Bolus administration of ALLO (0.07 mg/kg) in healthy 
women produces a small deterioration in verbal memory but has no effect on semantic and working 
memory [37]. Finally, bolus administration of a low dose of ALLO (0.05 mg/kg) in healthy women 
and women with PMDD results in mild sedation and no effect on startle response [38]. 
 


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Laboratory stress paradigms can predict outcome and relapse in AUD: Clinical 
observations and laboratory studies – including work done by our group (see preliminary data) - show 
that the stress response is strongly linked to alcohol craving, relapse, and increased alcohol intake [6, 
41]. Laboratory studies show that stress cues increase alcohol craving in active drinkers, abstinent 
alcoholics, and social drinkers[6, 41-44]. Laboratory studies using both stress inducing cues (fear, 
anxiety)[2, 45-49], and alcohol cues (smell of alcohol)[3, 50] show that stress cues are as powerful in 
eliciting self-reported craving for alcohol as is exposure to alcohol cues[48, 51, 52]. Also, stress-
induced craving predicts drinking outcomes in alcoholics followed for 90 days after discharge from an 
inpatient treatment program[2]. Individuals with greater stress reactivity in the lab have a shorter 
time to relapse to their preferred substance than individuals with lesser stress reactivity[53]. This 
paradigm has been used to test medications (such as cloninide, guanfacine, prazosin) for their 
potential to attenuate the stress response in the laboratory prior to testing in clinical trials. The 
laboratory stress paradigm we are testing has been successfully used by our group (preliminary 
data section) and others[42, 54], is a clinically relevant paradigm because of its potential to explore 
the link between reactivity to stress and alcohol use, and can be used to test underlying 
neurobiology and potential therapeutic agents. 
 
Proposed study: This project will investigate if pretreatment with the neurosteroid ALLO 
administered intravenously attenuates stress-induced craving, anxiety and subjective 
stimulant/sedative alcohol effects in those with AUD using a laboratory paradigm. Subjects (N=60 
completers) will receive a continuous infusion of ALLO to achieve a targeted plasma level of 100nM or 
placebo over 200 minutes. On a single test day, after 60 min of ALLO or placebo infusion all 
participants will undergo laboratory testing for stress-induced craving using personalized scripts 
based on an index stressful life event for each subject. This technique has been shown to produce 
robust and reliable craving and anxiety in subjects including heavy drinkers[1], alcohol dependent 
individuals[2] both with and without psychiatric comorbidity (including work by our group) and, in 
those with other substance use disorders[3-5]. All participants will also receive alcohol administered 
intravenously using a clamp procedure, targeting a breath alcohol concentration (BrAc) of 40mg% (40 
mg/dL). This procedure will achieve steady-state blood alcohol levels without the individual variation 
from oral alcohol administration. IV alcohol administration will start after the script presentation; it 
will be infused over 20 min. and clamped for an additional 30 min. ALLO and placebo will be 
identical in color, clarity and viscosity and the blind will be maintained. The collaborator (Rogawski) 
in this study has extensive experience administering ALLO infusions in individuals with seizure 
disorders [55, 56], and TBI (NCT01673828). In doses proposed in this study, ALLO is very well 
tolerated with mild sedation being the main side effect. 
 
INNOVATION: This study is among the first to evaluate administration of the neurosteroid ALLO 
for its effect on craving, anxiety, and subjective stimulant/sedative alcohol effects. Because of its CNS 
effects and its role in stress, it has a therapeutic potential for those with AUD and comorbidity, 
particularly PTSD and TBI. The investigators in this study represent a unique collaboration and 
together are ideally suited to conduct this research. Specifically, we have included experts on 
neurosteroids and the administration of ALLO (Rogawski), those who have expertise in alcohol 
dependence treatment (Ralevski, Petrakis), neurobiology and laboratory paradigms (Petrakis, 
Ralevski), and alcohol administration (Petrakis, Ralevski). Further, the investigators in this study 
have extensive expertise in the comorbidity of AUD with PTSD (Petrakis), and as such are well suited 
to further study this agent in this patient population. Finally, the study will carefully monitor the 
safety and effects of ALLO administration alone and in combination with alcohol in AUD, including 
its subjective effects, its effects on cognition, and on motor coordination.   
 
APPROACH 


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A. Our experience 
with laboratory stress-
induced paradigms: In 
a recent, randomized, 
within-subjects laboratory 
study, we compared the 
effects of neutral and 
stress cues (presented 
before [pre], and two times following the cues [post and recovery]) on craving and anxiety in 32 
individuals diagnosed with DSM-IV alcohol dependence (AD).  Fig. 1 and Fig 2 show the findings 
from the main outcome measures (Craving and anxiety measured by visual analog scales VAS) and 
indicate that stress when compared to neutral cues significantly increased alcohol craving and 
anxiety. The study shows that alcohol craving and anxiety can be reliably induced in the laboratory 
using this paradigm.   
 
B. Experience with ALLO administration: We have established a collaborative relationship with 
Dr. Rogawski whose expertise include administration of ALLO to patients with TBI (see Fig. 3), 
Alzheimer’s disease, and adults and children with seizure disorders [55, 56]. In this ongoing double-
blind, placebo-controlled, randomized, dose-finding, two-stage adaptive clinical trial, two doses of 
ALLO are compared to placebo when administered intravenously for 5 days beginning within 8 hours 
after injury in patients with moderate to severe brain injury. Treatments are administered during a 4-
day treatment period followed by a 1-day dose de-escalation period. Stage 1 of the study assesses 
safety and confirms the dosing plasma concentrations. Stage 2 allocates subjects to the 3 arms of 
treatment. Fig. 3 shows Stage 1 data for the low dose of ALLO in 6 participants. The primary outcome 
measure is The Glasgow Outcome Scale, 
administered 3 months after treatment to 
evaluate how the injury has affected functioning 
in major areas of patient life. Available data is 
currently not large enough for statistical 
analysis. 
 
C. Our experience with alcohol clamp: In 
previous studies, we have used the clamp 
method to safely administer alcohol [57, 58] to 
over 182 subjects. The clamp method refers to a 
procedure where alcohol is infused 
intravenously over a period of about 20 minutes 
and after that the infusion is “clamped” or 
maintained at the same level for a set period of time. We have used the clamp to evaluate differences 
in subjective effects of alcohol in healthy subjects with and without a family history of alcoholism [59], 
to evaluate genetic bases for differences in subjective effects in healthy subjects [60], and to assess 
alcohol’s effects on pain tolerance in healthy controls [57]. More recently, we demonstrated that IV 
nicotine reverses sedation and intoxication induced by IV alcohol in healthy volunteers [58]. This 
paradigm is ideally suited as a first step in testing a potential new medication for AUD as it allows the 
testing of subjective effects at a steady state of alcohol, without confounds of dose that are present 
when alcohol is orally administered.  
 
5. Research Plan:   
 
-1
1
3
5
7
Neutral
Stress
Fig. 1 VAS craving 
 
    Fig. 2 VAS anxiety  
-1
1
3
5
7
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Stress


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Overview: This is a double-blind, placebo-controlled, between-subjects study with 1 test day. 
Subjects will receive intravenous ALLO infusion to achieve a targeted plasma (100 nM; N=30, n=15 
men, 15 women) or placebo (PLA; N=30, n=15 men, n=15 women) in a randomized fashion.  We 
recognize that the double-blind in the study may be compromised by the lack of an active placebo. 
However, ALLO side effects or their intensity have not been extensively studied at this dose. The dose 
of alcohol is also so low that possible side effects of alcohol are also very slim. In addition, our group 
has extensive experience conducting similar studies where, for example, high dose of alcohol 
(significantly higher than in this study) is compared to a placebo. In order to protect the blind for the 
participants we have combined the possible side effects for ALLO, alcohol and placebo.    For each 
subject, the rate of infusion will be calculated based on their weight. After 60 min. of infusion, 
participants will be exposed to 2 conditions in random order: stress cues and neutral cues. The cues 
will consist of a 5 min. presentation of the stimulus (stress or neutral) followed by immediate 
evaluation of craving and anxiety. There will be a relaxation procedure between each condition (see 
Table in Human Subjects section), and the presentation of the second cue will be initiated after stress 
levels have decreased to baseline. During the first hour of infusion and before the start of the stress 
paradigm, all participants will complete a number of measures to characterize ALLO’s effects on 
mood, cognitive performance and motor coordination.  Following the script presentations all 
participants will receive IV alcohol (40 mg%) that will be infused over 20 min and clamped at a steady 
level for an additional 30 min (Study procedures are also presented in tabular form in the Human 
Subjects section). All subjects will make 2 visits, one to develop the scripts and one for testing. In 
order to control for baseline differences in ALLO we will study women in their follicular phase of the 
menstrual cycle when levels of ALLO are most stable. In the follicular phase, women have low ALLO 
levels that are comparable to those in men, < 1 nmol/L [61]. Women have higher ALLO levels than 
men in the luteal phase (> 4 nmol/L), and especially high during pregnancy (100 to 160 nmol/L)[62]. 
For females, all sessions will be scheduled within the first 4-7 days of a woman’s menstrual cycle. 
 
Please note: Since this is the first study that combines ALLO with alcohol we propose to run at least 
one subject under open-label condition at a targeted plasma of 50nM (To target 50 nM:  95.5 
mcg/kg/hr x 1 hour loading dose, then 28.7 mcg/kg/hr maintenance infusion, and at least one subject 
under open-label condition at a targeted plasma of 100Nm (To target 100 nM: 191 mcg/kg/hr x 1 hour 
loading dose, then 57.4 mcg/kg/hr maintenance infusion).  
 
Justification for use of ALLO and dose:  ALLO was chosen because it can be administered 
directly, and for its robust action on the GABAa receptors; this is believed to be responsible for the 
anxiolytic, analgesic, antiepileptic and neuroprotective properties of neurosteroids. We considered 
other neurosteroids, such as PROG. PROG is not of interest in this study because it does not bind 
directly to GABAa receptors and it has been widely accepted that the non-reproductive actions of 
PROG are mediated by ALLO. ALLO is currently not widely available in the US, but it has been 
studied previously in Europe. Dr. Rogawski’s laboratory has extensive experience with ALLO, and is 
the only laboratory that holds an IND for the clinical use of ALLO in the US. He will be providing 
ALLO for this study.   Dr. Ralevski will hold the IND (140598) for this study and will reference Dr. 
Rogawski’s IND for ALLO (IND111085) and Dr. Petrakis’ IND for Alcohol (IND121915).  However, the 
sterile injectable vials of ALLO will be compounded at an FDA registered 503B outsourcing facility. 
His lab is now investigating the efficacy of ALLO in TBI (NCT01673828) and Alzheimer’s disease 
(NCT02221622). We selected the dose in consultation with Dr. Rogawski, and is designed to 
maximize behavioral effects while limiting side effects such as sedation, alcohol-like intoxication, mild 
nausea, and flushing.  
 
Justification for study design: 1. We decided to use continuous infusion of ALLO because bolus 
(i.e., rapid IV push) infusion leads to rapid and dramatic increases in ALLO followed by a precipitous 


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drop (in about 20-40 min.). The approach of delivering the dose continuously with an infusion pump 
over the 200 min period will still result in fluctuating plasma levels but the fluctuations will not be as 
precipitous as those using bolus infusion. 2. The high cost of ALLO ($4175 per subject) combined 
with limited funding restricted our design to a single dose of ALLO, although a dose-ranging strategy 
would have generated important results regarding ALLO’s dose-ranging efficacy. 3. Personalized 
scripts were selected for stress induction rather than other stressors (Trier Social Stress test, or Cold 
Pressor Test) because they most reliably invoke stress in the lab [63], and they have been used to 
induce drug and alcohol craving. 4. We decided to administer both scripts in one laboratory session 
based on the review of the evidence from laboratory studies in addition to practical considerations.  
The proposed design has been successfully used by our group and others in numerous imaging studies 
with healthy subjects and individuals with Axis I disorders. The current design will also limit subject 
drop outs by significantly reducing the number of lab sessions. 5. Because the same stress condition 
cannot be administered on more than one occasion, a within subject design is not feasible for this 
study.  
 
Justification for inclusion/exclusion criteria: 1. We plan to enroll both men and women since 
ALLO has been safely given to both genders. We believe that gender limitations will make recruitment 
very difficult, and including both genders will make the study more generalizable. Also, since there are 
some indications that healthy men may be more sensitive to the effects of ALLO infusion than healthy 
women, it seems imperative to further elucidate possible gender differences by including both genders 
in the study design. 2. We are including smokers because of very high rates of smoking among heavy 
drinkers. In order to control for possible withdrawal during the lab session, all smokers will be 
allowed to have a cigarette before the beginning of the laboratory session.   
 
Justification for measures of specific aims: We selected the primary measures of craving and 
anxiety based on work using stress induction in the lab with AUD subjects. The selection of measures 
for subjective stimulant /sedative effects of alcohol was based on numerous studies using IV clamp 
procedures. The selection of measures to characterize the subjective/ behavioral effects of ALLO in 
this population was guided by documented effects of ALLO in healthy subjects and on well-
characterized effects of GABAa agonists (such as ethanol or benzodiazepines) in various populations.  
 
A. Human subjects: Subjects will be recruited from the New Haven area by newspaper 
advertisements, Craigslist, online postings through Trialfacts/Rewards, and flyers. Based on previous 
human laboratory studies, approximately 10 percent of the subjects are expected to either drop out or 
be terminated because of non-compliance with the study procedures. To meet the target sample of 60 
completers, about n=200 will be consented. After the initial phone screening, potential subjects will 
undergo a comprehensive evaluation that is summarized in Table 1. The administration of alcohol to 
human subjects as proposed in this study is in compliance with NIAAA guidelines for alcohol 
administration.  
 
TABLE 1. Assessments 
 
 
 
Study Procedures 
Screening/  
Baseline 
Test 
Day  
Follow up 
(1 Day 
and 4 
Weeks 
after Test 
Day) 
SCREENING & EVALUATION 
 
 
 
Screening Form 
X 
 
 
CIWA-Ar 
X 
X 
 


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Structured Clinical Interview for DSM-5 
(SCID) 
X 
 
 
EKG 
X 
 
 
Physical and Laboratory Examination 
X 
 
 
Vital Signs, Blood/Urine Pregnancy, 
Toxicology, BrAC 
X 
X 
 
Timeline Follow-Back (TLFB) 
X 
X 
X 
Script development 
X 
 
 
LABORATORY ASSESSMENTS 
 
 
 
Alcohol Urge Questionnaire (AUQ) 
 
X 
 
State Trait Anxiety Inventory (STAI-6) 
 
X 
 
Visual Analogue Scale (VAS)  
 
X 
 
Number of Drinks Scale (NDS) 
 
X 
 
Biphasic Alcohol Affects Scale (BAES) 
 
X 
 
Differential Emotions Scale (DES-R) 
 
X 
 
Addiction Research Center Inventory (ARCI) 
 
X 
 
Similarity to Drugs of Abuse Scale (SDAS) 
 
X 
 
Hopkins Verbal Learning Test-Revised 
(HVLT-R) 
 
X 
 
Go No-Go task 
 
X 
 
Grooved Pegboard Test 
 
X 
 
Plasma ALLO 
 
X 
 
Follow-up Survey 
 
 
X 
Inclusion criteria: 
1) Males and females, between the ages of 21 and 65; 2) Non-treatment seeking individuals with 
current DSM-5 AUD (please note we will exclude those with physiologic dependence on alcohol). This 
will be determined by the study physician conducting the physical examination, and by the Clinical 
Institute Withdrawal Assessment for Alcohol Scale (CIWA-Ar)[64] with those with score of ≥8 
excluded;  3) No current substance use disorder (except tobacco, alcohol, and marijuana); 4) No 
current medical problems and normal ECG; 5) For women, not pregnant as determined by pregnancy 
screening, not breast feeding.   
 
Exclusion criteria:  
1) Current major psychiatric illnesses including mood, psychotic, or anxiety disorders; 2) History of 
major medical illnesses; including liver diseases, heart disease, chronic pain or other medical 
conditions that the physician investigator deems contraindicated for the subject to be in the study; 3) 
Liver function tests (ALT or AST) greater than 3 times normal; 4) weight >120kg; 5) renal 
impairment; and 6) patients on the following medications: a) medications for alcoholism (e.g. 
naltrexone, disulfiram, topiramate, acamprosate); b) psychotropic medications that promote sedation 
(please note patients on psychotropic medications for current psychiatric conditions will also be 
excluded); and c) patients currently taking antibiotics or antifungals. 
 
Screening Labs: 
EKG, CBC w/differential, Chem7, LFTs, Calcium Profile, Amylase, Lipase, Free T4, TSH, GGT, 
Cholesterol (including LDH), PT/PTT/INR, urine toxicology, and urinalysis (routine and 
microscopic). Females only: beta HCG (blood, quantitative). 
 
B. Assessments: The assessments that will be used in this study are summarized in Table 1.  


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Main outcome measures: Craving for alcohol will be assessed using the Alcohol Urge Questionnaire 
(AUQ), 8 items measured on a 7-point Likert scale. Anxiety will be assessed using the State Trait 
Anxiety Inventory (STAI-6) – 6 item measure designed to assess trait and state aspects of anxiety. 
Sedation during the stress paradigm will be measured using a Visual Analogue Scale (VAS) (0=not 
sleepy at all to 100=falling asleep). Biphasic Alcohol Effects Scale (BAES) [65] is a 14-item self-
report adjective rating scale that will be used to measure the stimulant and sedative effects of alcohol.  
Cognitive functioning: Little is known about the effects of ALLO on cognitive functioning. However, 
GABA receptor agonists have been studied extensively and are known to negatively affect cognition. 
The effects of ALLO on various aspects of memory have been studied in animals and the data show 
that ALLO impairs spatial, working, and long-term memory [66, 67].  Only one study examined the 
effects of ALLO administration in healthy women and found that 0.07 mg/kg of ALLO had a 
statistically significant but relatively minor effect on verbal memory but no effect on semantic or 
working memory [37]. Verbal memory will be evaluated using The Hopkins Verbal Learning 
Test-Revised (HVLT-R). The HVLT-R is a word list learning test of verbal memory. The main 
outcomes for the HVLT-R are percent corrected for immediate and delayed recalls. GABAa agonists 
have also been shown to affect response inhibition or the ability to suppress a response in rare and 
irregular situations[68, 69]. We will use the Go No-Go task to evaluate response inhibition [70]. A 
series of rectangles are presented every 1150 ms and participants are instructed to press a spacebar 
when they see a green rectangle (=go) but refrain from pressing the spacebar when they see a blue 
rectangle (=no go), and to give equal importance to speed and accuracy. The primary outcome is the 
number of commission errors. Motor Coordination: The negative effects of GABAa agonists on motor 
coordination have been well documented and studied for decades[71]. Motor coordination will be 
assessed using the Grooved Pegboard Test (Lafayette Instrument Company). This test is a 
manipulative dexterity test, consisting of a board with randomly positioned slots in which subjects 
insert pegs. It is an eye-to-hand coordination test that has been used in research studies, including 
pharmacotherapy studies [72]. 
Plasma ALLO: Deidentified plasma levels of ALLO will be analyzed by Dr. Rogawski’s lab at UCDavis. 
Plasma samples will be obtained on multiple time points during the lab session (see Table in Human 
Subjects section) to examine how levels of ALLO relate to: a) mood, cognitive performance and motor 
coordination, and b) stress-induced craving and anxiety. 
 
Clinician Administered Assessments:  
 
Structured Clinical Interview for DSM-IV (SCID) [73]: Assessment used to determine 
psychiatric diagnoses.  This interview assesses DSM-IV current and lifetime psychiatric diagnoses for 
anxiety, mood, psychotic, alcohol and substance use, somatoform, and eating disorders.    
 
Time-Line Follow-Back Assessment Method (TLFB)): [74] Interview technique that will be 
used to obtain quantity/frequency of alcohol consumption data for each day during the 90-day period 
prior to the study, throughout the period of study participation (measuring drinking on days outside 
of test session day) and the follow-up.  Participants are given a blank calendar covering the time 
interval to be re-constructed and are asked to reconstruct retrospectively their drinking behavior over 
that interval.  The process is facilitated by establishing anchor points (e.g., holidays, anniversaries, 
major national events, etc.).  It can be scored to provide the number of days on which various levels of 
consumption occurred.  The time-line method has good test-retest reliability and good validity for 
verifiable events.  It has been used in numerous studies to compare pre- to post-treatment drinking. 
 
Self-Rated Craving Measures:  
 


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Alcohol Urge Questionnaire (AUQ) [75]: The AUQ is an 8 item questionnaire, derived from a 
larger 49 item "Questionnaire of Alcohol Urges," that assesses desire for a drink, expectation of 
positive effect from drinking, and inability to avoid drinking if alcohol was available.  The AUQ is a 
reliable and valid scale for the measurement of self-reported alcohol urges, and scores have been 
shown to be strongly related to alcohol dependence severity (as measured by ADS scores) and to 
cognitive preoccupation with alcohol.  Its brevity and time frame for ratings (i.e., right now) makes it 
suitable for administration during the alcohol infusion period.  
 
Biphasic Alcohol Effects Scale (BAES) [65]: The BAES is a 14-item self-report adjective rating 
scale that will be used to measure the stimulant and sedative effects of alcohol during the test 
sessions. This instrument has been found to be a sensitive and reliable measure to study medication 
influences on alcohol effects [76-78].   
 
Visual Analog Scales (VAS): In addition to sedation, the VAS has the following items that will 
also be assessed: craving for alcohol, high, anxious, drowsy, irritable, and nauseous.  We recognize 
that single item scales may have greater variability than multiple item scales.  However, they present a 
significant time saving advantage during busy test days.  Many of these subjective measures have been 
utilized in several previous studies at our Center where they show very good sensitivity to drug effects 
and convergent validity with other measures of these mood states and symptoms. 
 
State Trait Anxiety Inventory (STAI-6): The STAI-6 is a 6 item measure designed to assess trait 
and state aspects of anxiety. 
 
C. Study Procedures: All participants will be asked to arrive alcohol and drug free (except for 
marijuana) for the test session. Urine tests and a breathalyzer will be performed for verification. 
Those with positive urines or breathalyzer will be rescheduled one time. Additional non-compliance 
with this or any other procedure will result in discharge from the study. Since women will be recruited 
during the follicular phase of their menstrual cycle, they will be asked to call within the few days 
following the start of their menstrual bleeding to be scheduled for the laboratory session.  
 
Screening: Potential participants responding to advertisements will be given a brief description of the 
study over the phone and will then be asked to complete a phone interview to determine their 
preliminary eligibility to participate in the study.  If the caller appears to meet the eligibility criteria, 
he/she will be invited in for a 3 hour (approximate) screening session.*  During the screening visit, a 
detailed assessment will be completed to assess current health. Participants will be asked specifically 
about alcohol and drug use, and any traumatic experiences that they may have had. In addition, we 
will gather information about medical history, do an electrocardiogram (EKG), blood work (including 
pregnancy test for women), and urine will be tested for drugs of abuse (cocaine, marijuana, 
benzodiazepines, etc.).  This information will become part of the participant’s VA medical record.  The 
screening will last approximately 2-3 hours.  
 
*In order to reduce face-to-face interactions during COVID-19 potential participants 
will be given the option to complete a portion of the screening procedures remotely 
using a VA approved video platform (or by telephone when video is not feasible).   If the 
psychological screening is conducted remotely, the participant will be mailed/emailed a 
blank consent form and an informed consent process with study staff will then take 
place either over the phone or via VA-approved video platform.  We will go over the 
consent form in detail with the participants.  Consents that are mailed will be sent 
using US Mail (USPS/UPS/FedEx).  Consents that are e-mailed will be encrypted using a 
VA approved encryption platform (such as Azure).     


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A fully signed consent and HIPAA will be obtained prior to starting any screening 
procedures (remote or in-person) should they still be interested and eligible to 
continue in the study.  We will accept the following forms of signed consent and HIPAA: 
a) signed in person; b) signed at home and mailed back to the research team using a 
pre-addressed envelope; c) signed at home and the patient will take a photo of the 
consent and HIPAA pages and email the images to a member of the research team; or d) 
signed at home during a video visit where we will ask the patient to hold each consent 
and HIPAA page up to the camera and we will take a screenshot of the signature pages 
(ensuring the patient’s face is not in view) and print and file the screenshots in the 
patient’s research record.    
• The following assessments will NOT be conducted remotely: physical exam, blood 
pressure/pulse, blood work, urine toxicology, pregnancy test, and EKG.   
 
 
Randomization: Stratified block randomization will be used to balance treatment assignments within 
each gender.  Our group has used this procedure with repeated success in previous and ongoing 
studies. The Research Pharmacy will create the randomization tables. 
 
Script Development: Two scripts will be developed based on a procedure developed by Lang et al [79-
81]and using Scene Construction Questionnaires [81-83]. The questionnaires elicit details on the 
event and individuals involved, including physical sensations, thoughts, emotions, and cues related to 
the event described. The stress related scripts will be based on the “most stressful” recent experience.  
Only stressful experiences rated 8 and above will be used for script development. Stressful 
experiences related to substance use will not be considered. The neutral script will be based on 
previously developed neutral scripts that will consist of a relaxed beach scene commonly experienced 
by most individuals but personalized for each participant. The script development can also be done 
virtually. 
 
Alcohol Clamp Procedure: In this study, we will be using a modified alcohol-IV clamp procedure 
developed and standardized by Subramanian and colleagues [84], that has been used previously and 
in ongoing studies by the Co-PI [59]. We will use the latest state-of-the art infusion method CAIS that 
uses a computerized control of the alcohol infusion that includes real-time pharmacokinetic modeling 
to optimize the reliability and standardization of the procedure [85]. The infusion will be performed 
using a 6% ethanol solution in 0.9% saline. The computer assisted administration program 
automatically calculates and corrects the infusion rate based on real-time BrAc data entry by staff, 
based on the pharmacokinetic profile of each subject.  
 
Alternatively, the alcohol-IV clamp procedure may be completed by using a MATLAB [86] calculation 
package.  The loading phase rate, determined by MATLAB, will use a formula that includes patient 
age, gender, height, and weight to generate a linear ascension to target BrAc in 30 minutes.  The 
infusion will still be performed using a 6% ethanol solution in 0.9% saline.  BrAc will be measured 
during the loading phase as well as during the target BrAc clamp.  Once the target BrAc is achieved, 
the infusion pump rate will be adjusted so that the subjects are maintained within ±5 mg% of target 
BrAc for 30 minutes.  In previous studies, this alcohol clamp method has been used safely and 
successfully to administer alcohol [57-60] in over 182 subjects. 
 
Participants will be required to have a negative COVID test within 3 days of the Test 
Day. Participants who can show proof (completed vaccination card) that they are 
fully vaccinated will not be required to have a COVID test prior to any of the test days, 


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unless local guidelines (e.g., Yale University, VACT, the state of CT) require it at the 
time of study participation. 
 
After the test day is completed, subjects will be asked to remain on the Biostudies Unit to make sure 
that there are no after effects of the alcohol or ALLO. A physician will examine them before they are 
permitted to go home. After the test day, subjects will be encouraged to make arrangements to be 
driven home.  If they are driving themselves home, they will not be permitted to leave until their 
breathalyzer is at or below (≤) 0.02 and prior to discharge a physician will evaluate them to detect 
any signs of intoxication that would impair driving ability.   
 
Blood Draw: In addition to collecting ALLO, we will also be collecting Ghrelin and Neuropeptide Y 
(NPY).  Recent evidence suggests that peptides such as Ghrelin and NPY may play a role in stress, 
alcohol craving, and reward; however, this research is in its early stages. The role of these peptides in 
alcohol consumption and in alcohol use disorders is not well understood. Current evidence from both 
animal and human research demonstrates that the peptide Ghrelin is strongly and positively related 
to craving for alcohol. The role of NPY in relation to craving for alcohol is less understood. This 
project will attempt to understand the role of Ghrelin and NPY in drinking, stress, and craving in 
heavy drinkers. We will collect Ghrelin and NPY on a total of N=60 completers.  
 
Follow-up: After the test day, participants will be contacted for 2 telephone follow-ups. The first 
telephone interview will be approximately 1 business day after the participant completes their lab 
session. They will be asked about how they have been doing since their last test day as well as about 
any alcohol, tobacco, or drugs they may have consumed. The questionnaire will take about 10 minutes 
to complete. The week 4 follow-up will also be over the phone and will last about 30-60 minutes, 
where they will be asked about how they have been doing since the test day and will complete a TLFB 
for the past 4 weeks as well as other surveys outlined in Table 1. 
 
D. Data analysis and sample size considerations:   
Study data will be collected and managed using REDCap electronic data capture tools hosted at VA CT 
Healthcare System. REDCap (Research Electronic Data Capture) is a secure, web-based application 
designed to support data capture for research studies, providing 1) an intuitive interface for validated 
data entry; 2) audit trails for tracking data manipulation and export procedures; 3) automated export 
procedures for seamless data downloads to common statistical packages; and 4) procedures for 
importing data from external sources (Harris et al 2009). 
 
Data Analysis: Descriptive statistics will be calculated and normality assumptions will be checked 
prior to analyses. Transformations or non-parametric methods will be used if normality assumptions 
are not satisfied. The primary analyses will use linear mixed models (LMMs). LMMs are flexible in 
modeling the variance-covariance structure of the repeated measures, use all available data on an 
individual, and provide unbiased and efficient results when data are missing at random. Based on 
Schwartz Bayesian information criterion (BIC) (smaller is better), we will select the appropriate 
correlation structure for each dependent variable. Medication will be entered as a categorical variable 
with two levels (placebo, ALLO 100 nM). Stress/Neutral cues will be entered as a within-subject 
variable with two levels: stress and neutral. Time will also be a within-subject factor (before cue 
presentation, and 2 times after each cue presentation). Specific Primary Aims will examine the 
effect of medication on craving (assessed with AUQ), and anxiety (assessed with STAI-6) under stress 
vs. neutral condition, and stimulant/sedative effects of 40 mg% alcohol. We will consider a significant 
interaction between medication and stress condition with smaller differences between the stress and 
neutral conditions on active drug (ALLO) supportive of our hypothesis. Specific Secondary Aims 
will examine the effect of medication, under stress vs. neutral, and in combination with alcohol on 


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subjective mood effects, cognitive functioning and motor coordination. As stated in the specific 
primary aims, a significant interaction between medication and stress condition with smaller 
differences between the stress and neutral conditions on active drug (ALLO) vs. placebo will be 
considered supportive of our hypothesis. Exploratory Aims will examine the gender-specific effects 
by adding gender as a between-subject factor in the models above and testing all possible interactions 
involving gender. All statistical testing for the primary hypotheses will be two-sided at 0.05 
significance level. Post-hoc tests and analyses of secondary outcome measures will be adjusted for 
multiplicity using the Bonferroni correction.  
 
Sample size considerations: With 30 subjects per treatment group, we have at least 80% power to 
detect large effects (f=0.5) for the interactions of interest for Aims 1a, 1b, and 1c assuming 
alpha=0.05. We can also detect large effects for the comparisons of ALLO dose to placebo (d= 0.74) 
under the same assumptions. The effects we are proposing are based on data from Timby et al. 2006 
and van Broekhoven et al 2007 who reported that ALLO administration had a large effect (d>0.8) on 
saccadic eye movement and sedation among healthy subjects.    
 
6. PROTECTION OF HUMAN SUBJECTS: 
 
Human subjects: Non-treatment seeking individuals with AUD will be recruited from the New Haven 
area by newspaper advertisements, Craigslist, and flyers. Based on previous human laboratory 
studies, approximately 10 percent of the subjects who enroll in the study, meet all inclusion/exclusion 
criteria and are randomized are expected to either drop out or be terminated because of non-
compliance with the study procedures. To meet the target sample of 60 completers, a total of 66 
participants will be randomized. After the initial phone screening, potential subjects will undergo a 
comprehensive evaluation that is summarized in Table 1 in “Research Strategy”. 
 
Study Procedures: Table 1 delineates the timeline for study procedures and measures that will be 
administered at each time point during the laboratory session. 
 
Table 2. Study Procedures (times are approximate) 
Time  
Measures and Events 
Baseline 
Urine and BAC check, HR/BP, CIWA, TLFB, AUQ, STAI-
6, VAS; BAES, NDS, DES-R, ARCI, SDAS, HVLT-R, Go 
no-Go, Grooved Pegboard Test, Ghrelin, NPY and ALLO 
levels, breakfast 
-62 min 
Ghrelin and NPY levels 
-60 min 
ALLO infusion starts 
-40 min 
HR/BP, AUQ, STAI-6, VAS; BAES, NDS, DES-R, ARCI, 
SDAS, HVLT-R, Go no-Go, Grooved Pegboard Test, 
ALLO levels 
-20 min 
HR/BP, ALLO levels 
-2 min 
Ghrelin and NPY levels 
Stress Reactivity Condition 1 (15 min break) 
0 min 
Condition 1. (Neutral or Stress) Baseline/Relaxation; BP 
& HR, AUQ, STAI-6, VAS   
+10 min  
Image period, BP & HR  
+15 min 
BP&HR, AUQ, STAI-6, VAS, Ghrelin and NPY levels  
+20 min 
Recovery period. AUQ, STAI-6, VAS 
+25 min 
BP&HR, AUQ, STAI-6, VAS, Ghrelin, NPY and ALLO 
levels 


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Stress Reactivity Condition 2 
+30 min 
Condition 2. (Neutral or Stress). Baseline/Relaxation as 
described above 
+40 min  
Image Period as described above 
+45 min 
BP&HR, AUQ, STAI-6, VAS, Ghrelin and NPY levels 
+50 min 
Recovery Period as described above 
+55 min 
BP&HR, AUQ, STAI-6, VAS, BAES, NDS, DES-R, ARCI, 
SDAS, Ghrelin, NPY, and ALLO levels 
Alcohol Infusion 
+70 min 
start of IV alcohol infusion  
+90 min 
Target reached 
STAI-6, VAS, BAES, NDS, ARCI, SDAS, HVLT-R, Go no-
Go, Grooved Pegboard 
+118 min 
Ghrelin and NPY draw  
+120 min IV alcohol infusion ends, ALLO infusion ends  
+150 min STAI-6, VAS, BAES, NDS, ARCI, SDAS, HVLT-R, Go no-
Go, Grooved Pegboard, ALLO levels, Lunch 
6.1 Risk to the subjects  
Human subject’s involvement and characteristics:   
 
Sources of material: The subject will be the source of material. They will be informed that the research 
material collected will be for research purposes only. Blood, plasma, and urine samples will be 
obtained from study participants. The blood samples will be obtained with standard venipuncture 
techniques. In addition, physiological (heart rate and blood pressure), cognitive performance, and 
subjective measures will be obtained.  
 
Potential risks: There are potential risks, discomforts and inconveniences associated with the 
participation in this study. These may be due to allopregnanolone (ALLO) administration, IV alcohol 
administration, combination of ALLO and alcohol, stress induction, blood draw and loss of privacy. 
1) Allopregnanolone: In this study we will use a targeted plasma of 100 nM of ALLO. This dose of 
ALLO is similar to the levels found in women in their third trimester of pregnancy. Similar 
doses have been safely given to healthy participants with minimal and transient side effects. 
Common adverse effects associated with single administration of ALLO are sedation, alcohol-
like intoxication, headaches, mild nausea and flushing. Less common side effects associated 
with neurosteroids, such as progesterone, and reported after long-term use include depression, 
anxiety, blockage of blood vessels, and increased risk for heart attack or stroke. 
2) IV alcohol administration: Adverse effects resulting from ethanol consumption include blurred 
vision, nausea, vomiting, flushing, headache, and lightheadedness.  These side effects should 
reach their peak within 1.5 hours following consumption of alcohol and decline thereafter.   
3) Combined administration of ALLO and alcohol: We anticipate that ALLO will attenuate the 
sedative and stimulant effects of alcohol. However, since this will be the first instance the two 
compounds are given together all participants will be continuously monitored, a physician 
and/or a nurse will be present during the laboratory session at all times.   
4) Stress induction: The interview, questionnaires, and imagery sessions will require subjects to 
discuss stressful experiences. These procedures are expected to cause a moderate degree of 
anxiety, psychological discomfort, and alcohol craving that will return to baseline levels before 
the subjects are sent home.  If the subject continues to experience unpleasant symptoms a few 
hours after the end of the procedures, clinically-trained staff will guide the subject through 
relaxation techniques. If urges to drink or emotional distress persist after an hour of relaxation 
training, the subject will receive an individual counseling session with a psychologist who is 


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experienced in psychotherapy.  If urges to drink or emotional distress persist after the 
psychotherapy session, the subject will be escorted to the Emergency Department and treated 
by the on-call psychiatrist.   
5) Blood Drawing: Subjects will have approximately 140 ml of blood drawn as a result of their 
participation in the study. Blood drawing can cause some pain and may result in bruising. 
6) Loss of confidentiality: Participation in the study may lead to loss of confidentiality. 
 
6.2 Adequacy of protection against risks.  
General procedures to protect against risks: Before initiating any research activity, each subject must 
give informed consent that will detail the risks of study participation. Eligibility will be determined by 
the medical and psychiatric history, drug use history, and the physical examination done prior to 
beginning this research protocol. Subjects will be provided a number to call to reach an on-call 
psychiatrist (24 hours/day) should unpleasant effects occur after subjects have left the testing facility. 
We ask subjects to contact us on the day following tests on a routine basis and if we are not contacted, 
we will call the subjects to monitor their status. 
 
1) Protection against adverse events from ALLO: To minimize risks associated with ALLO, 
subjects will be closely monitored throughout the laboratory session. The targeted plasma level  
of ALLO (100 nM) used in this study has been used safely in studies with healthy human 
subjects. The dose is higher than circulating levels of ALLO in men and women but comparable 
to circulating levels recorded in women during pregnancy.  
 
2) a) The administration of alcohol to human subjects proposed in this study is in compliance 
with NIAAA guidelines for alcohol administration.  Subjects are carefully screened so only 
those who have experience with the dose of alcohol will be enrolled and those that are 
treatment seeking will be excluded. The clamping procedure is safe and has been used safely 
with blood alcohol levels up to 0.15 g/dL. The ethanol dose employed in this study is equivalent 
to doses used in previous studies in healthy human subjects and falls within ranges that alcohol 
dependent individuals self-administer. No fetus will be exposed to alcohol: pregnancy tests will 
be administered on the day of testing to every female subject with positive results excluding 
participation. Women who are breastfeeding will not be included in this study. 
b) We have instituted several levels of medical monitoring including: i) the testing facility is 
covered at all times by the VA Connecticut Healthcare System (VACHS) medical emergency 
(“Medical Code”) team, ii) the nurses in the testing facility are all certified in advanced cardiac 
life support, iii) there is continuous medical and nursing presence throughout the test day, iv) 
subjects will be administered alcohol only if the systolic blood pressure is <150 mmHg and 
heart rate is <90 beats/minute. Subjects will be terminated from the study if the blood 
pressure at any time is >170/110 mm Hg or if the heart rate is >130 beats/min.  
c) At the end of each test day, the research staff will review the experiences of the day with the 
research subject.  Subjects will not be released from the Laboratory until the BrAc is less than 
or equal to 0.02 g/dL (0.02 g/dL is below the threshold of BrAc associated with any 
impairment in locomotor coordination or judgment in humans) and research personnel have 
documented that participants exhibit an alert and oriented demeanor.    
d) At the end of the test session, we will provide participants with a brief motivational 
interview based on the principles of Miller’s Motivational Enhancement Therapy. This 
interview will be conducted by a member of our research group, an M.D. or Ph.D. previously 
trained in motivational interviewing. During this 30minute interview, participants’ drinking 
and smoking behavior (for the subgroup of smokers), and experience of alcohol and smoking-
related problems will be reviewed. Participants will be presented with normative drinking 
information, as well as guidelines for non-hazardous alcohol consumption. If interested, 


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participants will be given treatment referrals to address their drinking and smoking behavior 
(for smokers).  It has been found that similar brief advice is associated with decreases in 
alcohol drinking behavior and increased motivation to quit drinking. 
e) All participants will be contacted approximately 1 business day and 1 month after the end of 
their participation to ensure no residual effects from ALLO or alcohol administrations. 
 
3) Stress induction: Similarly to above we have instituted several levels of medical monitoring 
including: i) the testing facility is covered at all times by the VA Connecticut Healthcare System 
(VACHS) medical emergency (“Medical Code”) team, ii) the nurses in the testing facility are all 
certified in advanced cardiac life support and one of the four nurses have extensive medical 
ICU experience, iii) there is continuous medical and nursing presence throughout the test day, 
iv) at the end of the test day, the research staff will review the experiences of the day with the 
research subject. The subject will not be released from the unit unless his/her anxiety, 
psychological discomfort, and alcohol craving have returned to baseline levels.  At the end of 
the lab session, we will provide participants with a brief motivational interview based on the 
principles of Miller’s Motivational Enhancement Therapy (details outlined above).  
 
4) Blood drawing: The medical staff who perform blood drawing are trained individuals and 
follow Standard Precautions. 
 
 
5) Confidentiality will be protected by having records identified by code number only with the 
master list including names kept in a sealed envelope in a locked file in the Principal 
Investigator's office and by the pharmacy, or on the VA secure server. Subjects will be given 
telephone numbers to call in case of emergency, 24 hours a day.    
 
Stopping rules: Subjects will be terminated from the study if the blood pressure at any time is 
>170/110 mm Hg or if the heart rate is >130 beats/min. Subjects will remain in the laboratory until 
the breathalyzer reaches ≤ 0.02 g/dL. These procedures have been developed as part of our routine 
procedures for IV alcohol administration. Blood pressure and heart rate will be monitored frequently 
during the lab session.  If blood pressure and/or heart rate exceeds these parameters, the infusion will 
be terminated. If blood pressure does not improve after the infusion is terminated, an MD will 
evaluate the subject and make a determination regarding rescue medications.  If symptoms do not 
improve the patient will be escorted to the ER if deemed necessary. 
 
 
6.3 Potential benefits of the proposed research to the subjects and others. Study participation may 
help the participants to seek help for their alcohol use. At the end of the study, if interested, 
participants will be referred to clinical programs. 
 
6.4 Importance of the knowledge to be gained. This proposed study may help to develop new and 
more effective treatments for AUD. We believe that the risk/benefit ratio for this study is acceptable, 
and that the benefits of the proposed studies outweigh the potential risks to subjects.  
 
6.5 Data safety and monitoring plan. The risk associated with participating in this study is moderate, 
because ALLO administration may be associated with mild side effects and the effects of the stress-
induction procedure are expected to normalize quickly. Serious side effects associated with this study 
are not expected. This project will be monitored by a Data and Safety Monitoring Board (DSMB), 
because the study involves double-blind administration of ALLO in combination with alcohol in AUD 
participants. This board is composed of persons not otherwise affiliated with the study who are 


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experienced in various aspects of the conduct of clinical trials, laboratory studies and treatment of 
addictive disorders. We propose three investigators located here in Connecticut who are not directly 
involved in this study – Declan Barry, Ph.D., Sherry McKee, Ph.D., and David Fiellin, M.D. as the 
members of the DSMB. The members of the DSMB and all study Investigators will complete Conflict 
of Interest forms created by Yale’s IRB in accordance with NIH guidelines.   
 
We will report recruitment, follow-up, and adverse events to this panel on a quarterly fashion.  Prior 
to study initiation, critical parameters for collection of side effects and for study discontinuation will 
be recommended to the DSMB who may use these or other measures to monitor safety of the ongoing 
study. The DSMB will be available to convene outside of scheduled meetings, if necessary, due to 
concerns regarding a particular subject or due to any troublesome developments in subjects' 
experiences during the study. The DSMB will make appropriate recommendations for changes in the 
study protocol, if needed.   
 
This monitoring will be consistent with NIH policy regarding the protection of human subjects in 
research, and FDA guidance on statistical practices for clinical trials (ICH E9) and good clinical 
practices (ICH E6). In general, the data to be reviewed will include screening data, baseline data, 
laboratory data, and safety data.   
The Principal Investigator will conduct a review of all adverse events and determine the attribution 
and grade of severity of the adverse event by using the following scales: 
Attribution of Risk Categories: 
Definite:  
Adverse event(s) will clearly be related to investigational agent(s) or other intervention 
Probable: 
Adverse event(s) will likely be related to investigational agent(s) 
Possible:   
Adverse event(s) may be related to investigational agent(s) 
Unlikely:  
Adverse event(s) will doubtfully be related to investigational agent(s) 
Unrelated: Adverse event(s) will clearly not be related to the investigational agents(s) 
Grades of Risk: 
 
 
0: 
No adverse event or within normal limits 
 
 
1:  
Mild adverse event 
 
 
2:  
Moderate adverse event 
 
3:  
Severe adverse event resulting in hospitalization or prolongation of existing 
hospitalization, a persistent or significant disability/incapacity, or a congenital anomaly/birth defect  
 
 
4:  
Life-threatening or disabling adverse event 
 
 
5:  
Fatal adverse event 
Serious adverse events (SAEs) include any untoward medical occurrence that results in death or the 
immediate risk of death, hospitalization or the prolonging of an existing hospitalization, persistent or 
significant disability/incapacity or a congenital anomaly/birth defect. Subjects will be terminated 
from participation if the investigator feels that subjects' health or well-being may be threatened by 
continuation in the study. Serious unanticipated and anticipated adverse events will be reported 
within 48 hours to the VA Hospital and Yale IRBs, and NIAAA. We will directly report to the FDA, 
whenever their magnitude or frequency exceeds expectations.   
 
6.6 Clinicaltrials.gov requirements. This study will be registered at the clinicaltrials.gov website.  
  
7. Payment:  
Participants will be paid $30 for the screening session, $30 for the script development session, $150 
for attending the test session, and $0 for the 1 month follow-up. Thus, the total amount that could be 
earned during the study is $210.  This payment is for the time and effort associated with study 
assessments and procedures.  This payment will be given in the form of cash. Participants will not be 
paid if a test day is cancelled because they test positive for alcohol or illicit drugs in their system.  If a 


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patient participates in a virtual screening and is not eligible to come for the in-person visits they can 
choose to come in to pick up their cash payment or be mailed a gift card or check.  If they choose to 
receive payment by gift card or check they may need to complete a tax form. 
 
We will provide participants with 2 meals during the lab visit (breakfast and lunch). 


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8.  References:  
 
 
1. 
Ray, L.A., et al., A human laboratory study of the effects of quetiapine on subjective 
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Breese, G.R., et al., Stress enhancement of craving during sobriety: a risk for relapse. 
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Coffey, S.F., et al., Trauma and substance cue reactivity in individuals with comorbid 
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Coffey, S.F., et al., Trauma-focused imaginal exposure for individuals with comorbid 
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Timby, E., et al., Pharmacokinetic and behavioral effects of allopregnanolone in healthy 
women. Psychopharmacology (Berl), 2006. 186(3): p. 414-24. 


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ICF_000.pdf: 
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022 
RESEARCH SUMMARY
You are invited to take part in a research study because you are between 21 and 65 years of 
age, have alcohol use disorder (AUD), and are not seeking treatment. This study is sponsored 
by Yale University. Before you decide to take part, it is important for you to kinow why the 
research is being done and what it will involve. This initial summary is to give you key 
information to help you decide whether to participate. Detailed information follows this brief 
summary. Ask the research team questions. Taking part in this study is completely voluntary.
WHAT IS THE STUDY ABOUT AND HOW LONG WILL IT LAST?  
We are asking you to choose whether or not to volunteer for a research study about the effects 
of allopregnanolone (ALLO) on the subjective effects of alcohol. ALLO is a product of the 
hormone progesterone and is found naturally in your body. This initial summary is to give you 
key information to help you decide whether to participate. We have included detailed information 
after this brief summary. Taking part in this study is completely voluntary.
Your participation in this research will last about two months. 
If you decide to participate in this research study, you will be asked to complete a screening 
visit, script writing session, one test day,  and 2 telephone follow-ups, one approximately one 
business day after the test day, and another follow up one month later. On the test day, you will 
be given either an intravenous ALLO infusion or placebo (saline). You will have a 1 in 2 chance 
of being administered ALLO, much like the probability of flipping a coin. Everyone who 
participates in the test day will receive an intravenous infusion of alcohol.  Other study 
assessments include blood draws, urine drug and pregnancy tests, EKG, physical examination, 
interviews, questionnaires, and assessments. 
WHAT ARE KEY REASONS YOU MIGHT CHOOSE TO VOLUNTEER FOR THIS STUDY?
You have been invited because you are between 21 and 65 years of age, have an alcohol use 
disorder (AUD), and are not seeking treatment.  Although this study is not designed to benefit 
you personally, your participation may lead to knowledge that may help others.
WHAT ARE KEY REASONS YOU MIGHT CHOOSE NOT TO VOLUNTEER FOR THIS 
STUDY?
Administration of ALLO in humans is very novel with only a few studies being conducted to date 
and none in this population. The dose of ALLO selected for this study is similar to the levels 
found in women in their third trimester of pregnancy. Similar doses have been safely given to 
healthy participants in a small number of studies with minimal and transient side effects. 
You may experience side effects associated with alcohol/ALLO/placebo, including sedation, 
alcohol-like intoxication, blurred vision, headaches, lightheadedness, mild nausea, vomiting, and 
Page 1 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
NCT04015869           Protocol ID: 2000021601 


APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                 
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022
flushing. Less common side effects associated with neurosteroids, such as progesterone, and 
reported after long-term use include depression, anxiety, blockage of blood vessels, and 
increased risk for heart attack or stroke.  However, since this will be the first-time alcohol/ALLO/
placebo will be given together you may experience other side effects that have not been listed 
but a physician and/or a nurse will be present during the laboratory session at all times to 
continuously monitor you.  
The script development session will require you to discuss stressful experiences. These 
procedures are expected to cause a moderate degree of anxiety, psychological discomfort, and 
alcohol craving.  However, it is possible that you may experience a more intense reaction. 
Should you experience any unpleasant symptoms; clinically-trained research staff will guide you 
through relaxation techniques. One of the study physicians will be prepared to provide physical 
and medical assistance, and will be available for further evaluation, treatment, and follow-up. If 
necessary, you may be referred for further evaluation and/or treatment. For a complete 
descriptions of risks, refer to the Research Details.
You should not participate in this study if you are pregnant or currently breastfeeding.
DO YOU HAVE TO TAKE PART IN THE STUDY?
If you decide to take part in the study, it should be because you really want to volunteer. You will 
not lose any services, benefits or rights you would normally have if you choose not to volunteer. 
RESEARCH DETAILS
WHAT IS THE PURPOSE OF THIS STUDY?
With this research we hope to learn how ALLO, compared to placebo, influences how people 
experience the effects of alcohol, including craving, anxiety, and stimulant/sedative effects.  
ALLO is not approved by the FDA as a treatment for AUD. 
HOW LONG WILL YOU BE IN THE STUDY?
This research study is expected to take approximately 2 years. Your individual participation in 
the project will take approximately 2 months. Approximately 30 men and 30 women will 
complete the study.
WHAT WILL HAPPEN IF YOU TAKE PART IN THE STUDY?
Since ALLO is not an FDA approved treatment for AUD, this is an experimental study.  If you 
decide to participate in this research study, you will be asked to participate in the following:
 
Screening visit: 2-3-hour visit involving interviews, questionnaires, physical exam, EKG, 
blood and urine tests
Page 2 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022


APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                 
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022
 
Script writing session: 2-hour visit to complete a questionnaire and discuss the most 
stressful event you have experienced, creation of audio tape
 
Test day: 6-8-hour visit involving blood and urine tests, questionnaires, assessments, IV 
infusion of ALLO or placebo, tape session, IV infusion of alcohol 
 
Telephone follow-up 1: 10-minute follow up phone call 
 
Telephone follow up 2:  30-60-minute visit scheduled approximately 1-month after the 
test day to answer questions and a follow up survey.
WHAT IS EXPECTED OF YOU IF YOU TAKE PART IN THIS STUDY?
Screening and clinical assessments:
During the screening visit, a detailed assessment will be completed to assess your current 
health. You will be asked specifically about alcohol and drug use, and you will also be asked 
about any traumatic experiences that you may have had. In addition, we will gather information 
from your medical history and complete a physical assessment. As part of the physical 
assessment, an electrocardiogram (EKG), blood work, and drug test will be completed. Your 
urine will be tested for drugs of abuse (cocaine, marijuana, benzodiazepines, etc.); if you are 
female, a blood pregnancy test will also be performed.  If your urine is positive for drugs, except 
marijuana, or if you are pregnant, you will not be allowed to participate in the study.  All lab 
results will become part of your medical record at the VA. The screening will last approximately 
2-3 hours and it may take one week before we receive all the test results.  You will not be 
charged for these tests. At this point the physician in charge of the study will determine if you 
are eligible to participate. 
At any time during the screening evaluation, or study days, your participation may be terminated 
without your consent if we determine that you do not meet the entry criteria, or if the study 
physician feels it would be harmful for you to continue participating.
You will have the option to complete part of this screening visit remotely using a VA approved 
telephone or video platform.  If you choose this option a member of the research team will 
contact you to set up an appointment to go over this consent with you.  If you would still like to 
participate in the study after reviewing the consent, we will obtain verbal consent to continue 
and this will be documented in your medical record.  You will need to sign written consent at 
your first in-person study visit.
For female subjects of childbearing potential:
If you are a female subject you will have a blood pregnancy test done at screening, and you will 
have a urine pregnancy test when you come in for the test day.  Because alcohol and ALLO 
both have adverse effects on the fetus, you will not be able to participate in this study if you are 
pregnant or breastfeeding.  To your knowledge, you are not pregnant at this time.  You also 
agree to avoid becoming pregnant.  If you change your mind about becoming pregnant we ask 
that you tell us.
Page 3 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022


APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                 
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022
Script Development Session:
 
  During the script development session, you will be asked to recall 
the most stressful event that you have experienced. A questionnaire will be completed to gather 
specific information about this event. Stressful experiences related to substance use will not be 
considered. An audiotape of the event lasting approximately five minutes will then be created. 
We will also develop a neutral script, but this will be based on previously developed neutral 
scripts that will consist of a relaxed beach scene commonly experienced by most individuals but 
personalized for you.  This appointment will take approximately 2 hours and can be done in 
person or by a VA approved telephone or video platform.
Since levels of ALLO fluctuate in women we will ask all female participants to call our lab 
when their menses start. If you are a woman, you will be scheduled for your laboratory 
test session within the first 4-7 days of your menstrual cycle. 
Test Day: If you are eligible for the study on the basis of the screening and other clinical 
assessments, a member of the research team will contact you to schedule your test day, if you 
decide to participate. 
You will be required to have a negative COVID test within 3 days of the test session.    If 
you show proof that you are fully vaccinated you will not be required to have a COVID 
test prior to the test day, unless local guidelines (e.g., Yale University, VACT, the state of 
CT) require it at the time of your participation in the study.
For the test day, you will come to the Neurobiological Studies Unit (known as Biostudies), also 
located at the West Haven VA (Building 1, 9th floor) for testing.  The test day will start at 
approximately 7:30am and will last for 6 to 8 hours.  You will be asked to arrive with no 
alcohol or illicit drugs (except marijuana) in your system.  Since we will provide you with a 
light meal, we ask that you do not eat anything after midnight before coming in for the test 
day.  You can drink water before the test day.  If you smoke cigarettes you can smoke 
cigarettes as you normally do, until you come in for the test day; you will not be able to smoke 
cigarettes during the test day. 
When you arrive, the research nurse will take your blood pressure, pulse, and breath alcohol 
level and collect a urine sample that will be tested for illicit drugs. If you are a female, we will 
also do a urine pregnancy test.  If these tests indicate that you have recently used drugs (other 
than marijuana) or alcohol, or that you are pregnant, you may be discharged from the study.  If 
the study physician determines that it is unsafe for you to leave the hospital, you may be taken 
to the Psychiatric Emergency Room.
A thin plastic tube called an IV line will be inserted into a vein in each of your arms. Both IV lines 
are to permit administration of study drugs and for blood drawing. Blood will be drawn several 
times during the test day.  Both you and the research staff will not know if you were 
administered ALLO or a placebo (saline) for the duration of the time that you are involved in the 
study. We will also take your blood pressure, and your pulse will be monitored continuously. 
Page 4 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022


APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                 
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022
After the IV lines are placed, you will be asked some questions regarding the way you feel and 
think.  You will be asked similar questions throughout the study.  You will also be asked to take 
a computer test which involves watching a screen and pressing a button according to the 
instructions given to you.  After completing the initial questions, administration of either ALLO or 
a placebo (saline) will begin. 
You will then be presented with a tape that describes your most stressful event or decribes a 
pleasant, neutral event. You will hear the stress tape 1 time and the neutral tape 1 time in a 
random order.  You will then be asked to complete a variety of questionnaires and computer 
tasks, including ones asking about your alcohol craving, anxiety, mood, and cognitive 
functioning.  
If you experience unpleasant symptoms a few hours following tape presentation, a clinically-
trained staff member will guide you through relaxation techniques. If urges to drink or emotional 
distress persist after an hour of relaxation training, you will then receive an individual counseling 
session with a psychologist who is experienced in psychotherapy. If urges to drink or emotional 
distress persist after the psychotherapy session, you will be escorted to the Psychiatric 
Emergency Department and treated by the on-call psychiatrist.
Following the tape session, you will receive alcohol (breath alcohol concentration target of 40mg
%- about what is associated with approximately 2-12 oz. cans of beer in an hour for a man or 1-
2 12oz cans of beer for a woman) over a 50-minute period.  The alcohol will be administered 
through one of the IV lines that will be placed in your arm. Your breath alcohol concentration will 
be maintained by using a small pump that will regulate the flow of a solution containing alcohol 
into a vein in your arm.  The infusion pump rate will be adjusted every few minutes, based on 
the results of the breathalyzer tests.  On the test day, your breath alcohol concentration may be 
as high as 40mg% (Breathalyzer value of 0.04).    At several points during the alcohol 
administration you will complete a variety of questionnaires and computer tasks, including ones 
asking about your alcohol craving, anxiety, mood, and cognitive functioning.  You will also be 
asked to do these questionnaires several times after the alcohol administration is completed, 
until you are cleared to go home.
After the test day is completed, you will be asked to remain on the Biostudies Unit to make sure 
that there are no after effects of the alcohol or ALLO. A physician will examine you before you 
are permitted to go home. After the test day, you will be encouraged to make arrangements to 
be driven home.  If you are driving yourself home, you will not be permitted to leave until your 
breathalyzer is at or below (≤) 0.02 and prior to discharge a physician will evaluate you to detect 
any signs of intoxication that would impair your driving ability.  The option to drive yourself home 
is available only if there are no other methods of transportation available (e.g. public 
transportation, Yale shuttle, friends etc.).  This must also be approved by the Principal 
Investigator (Elizabeth Ralevski, PhD).  If you are deemed unsafe to leave due to intoxication, 
you may be kept against your will.  
 
Follow-up: After your test day is complete, you will be contacted approximately 1 business day 
and 1 month after your last test session for a telephone interview.  During the first telephone 
Page 5 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022


APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                 
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022
interview you will be asked about how you have been doing since the test day.  The phone 
interview will last approximately 10 minutes.  About four weeks after your test day is completed, 
you will be contacted again for a 30-60-minute follow-up interview, where you will be asked 
questions about your drinking and how you have been doing since the test day.
There is always a chance that any procedure can harm you.  The procedures in this study are 
no different.  In addition to the risks described above, you may experience a previously 
unknown risk or side effect. 
Risks of the usual care you receive are not risks of this study.  Those risks are not included in 
this consent form.  You should talk with your health care providers if you have any questions 
about the risks of usual care.  
WHAT POSSIBLE RISKS OR DISCOMFORTS MIGHT I HAVE IF YOU TAKE PART IN THIS 
STUDY?
Any procedure has possible risks and discomforts.  The procedures in this study may cause all, 
some, or none of the risks or side effects listed.  Rare, unknown, or unexpected risks also may 
occur.  
Effects of Alcohol/ALLO/placebo: Administration of alcohol/ALLO/placebo may cause alcohol-
like intoxication, blurred vision, sedation, headaches, lightheadedness, mild nausea, vomiting, 
and flushing. Less common side effects associated with neurosteroids, such as progesterone, 
and reported after long-term use include depression, anxiety, blockage of blood vessels, and 
increased risk for heart attack or stroke.  However, since this will be the first-time alcohol/ALLO/
placebo will be given together you may experience other side effects that have not been listed 
but a physician and/or a nurse will be present during the laboratory session at all times to 
continuously monitor you.  
To minimize possible injury as a result of alcohol consumption, you will be kept on the 
Biostudies Unit until your blood alcohol level is at a very low level (Breathalyzer at or below (≤) 
0.02).  For your protection, we encourage you to make arrangements to be driven home after 
the test day.  If you do not feel comfortable and don't want to continue or the research staff feels 
that you are at risk the test day will be discontinued.
Stress induction: The script development session will require you to discuss stressful 
experiences. These procedures are expected to cause a moderate degree of anxiety, 
psychological discomfort, and alcohol craving.  However, it is possible that you may experience 
a more intense reaction. Should you experience any unpleasant symptoms; clinically-trained 
research staff will guide you through relaxation techniques. One of the study physicians will be 
prepared to provide physical and medical assistance, and will be available for further evaluation, 
treatment, and follow-up. If necessary, you may be referred for further evaluation and/or 
treatment.
Page 6 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022


APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                 
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022
The Intravenous Lines and Blood Drawing: Blood drawing may cause pain, bruising, and 
rarely fainting or infection.  There is a chance of inflammation (swelling), bruising or a blood 
clot forming at the site of blood sampling.  Antiseptic technique will be used to minimize the 
risk of infection. Your blood will be drawn at your screening appointment and on your test 
day to provide information about your blood alcohol levels as well as other blood levels. You 
will have approximately 140 ml (5oz) of blood drawn over the course of this study and should 
refrain from blood donations eight weeks before and after testing.  
Study procedures: During the test day, you may not leave the test room. At the end of the test 
day, you will remain in the Biostudies Unit until we are satisfied that you are in good health and 
your Breathalyzer is at or below (≤) 0.02. During the study, you will be asked questions related 
to your mental health and drug use history which may make you feel uncomfortable, sad or 
bored. If you feel the questions are too stressful and would like to stop the screening or test day, 
you may do so at any time and your participation will be terminated.  In addition, you may be 
discontinued from the research study at any time if the study physician feels it is in your best 
interest to do so or if you are found to not be compliant with the study requirements (for 
example, if you tested positive for illicit drugs while enrolled in this study; if it becomes medically 
unsafe for you to participate; or, if the HSS withdraws approval).
Alcohol and ALLO both have adverse effects on the fetus, so you should not participate in this 
study if you are pregnant or breastfeeding.
There is always a chance that any procedure can harm you.  The procedures in this study are 
no different.  In addition to the risks described above, you may experience a previously 
unknown risk or side effect.  Risks of the usual care you receive are not risks of this study.  
Those risks are not included in this consent form.  You should talk with your health care 
providers if you have any questions about the risks of your usual care.  
WHAT ARE THE POSSIBLE BENEFITS OF THIS STUDY?
Taking part in this study is not designed to benefit you personally, but your participation may 
lead to knowledge that may help others.
HOW WILL YOUR PRIVATE INFORMATION BE PROTECTED?
Storage and Future Use of Data or Specimens:
Your information or biospecimens collected as part of the research, even if identifiers are 
removed, will not be used or distributed for future research studies.
Confidentiality of Information: Participation in research may involve a loss of privacy.  Your 
research records will be kept as confidential as possible.  Only a code number will identify your 
research records.  The code number will not be based on any information that could be used to 
Page 7 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022


APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                 
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022
identify you (for example, social security number, initials, birth date, etc.).  The master list linking 
names to code numbers will be kept separately from the research data.  
All research information will be secured in locked files.  Your identity will not be revealed in any 
reports or publications resulting from this study.  Only authorized persons will have access to 
the information gathered in this study.  Authorized persons may include regulatory agencies 
such as the Food and Drug Administration, (FDA), the Government Accountability Office (GAO), 
the Office for Human Research Protections (OHRP), Office of Research Oversight (ORO), VA 
Connecticut Healthcare System Research Office, and the Yale University Human Investigation 
Committee (HIC). Yale Accounts Payable will have limited access to your information (printed 
name and signature on your cash payment receipt).
The Department of Veterans Affairs (VA) requires some information to be recorded in the VA 
electronic medical record for veteran and non-veteran research subjects.  Therefore, if you 
participate in this study, a medical record will be created if you do not already have one.  Notes 
from your visits, procedures, and laboratory tests will be included in this record.  In addition to 
the research team, and the VA staff who provide clinical services, other researchers may be 
granted approval to access this information in the future.  Federal laws and regulation that 
protect privacy of medical records will apply to your VA record.  
There are exceptions to the promise of confidentiality.  For example, if we see or are told that a 
child is being abused or neglected, or that there is a risk of harm to you or others, we will 
disclose this information to the proper authorities.
Certificate of Confidentiality:
 We have obtained a Certificate of Confidentiality from the Federal Government.  This helps 
protect your privacy by allowing us to refuse to release your name or other information outside 
of the research study, even by a court order.  The Certificate of Confidentiality will not be used 
to prevent disclosures to local authorities of child abuse or neglect, or harm to self or others.  
The Certificate does not prevent you or a member of your family from releasing data about 
yourself or your involvement in this study.  
Medical Record 
We will include information about your study participation in your medical record.
Clinical Trial 
A description of this clinical trial will be available on http://www.ClinicalTrials.gov as required by 
U.S. Law.  This website will not include information that can identify you.  At most, the website 
will include a summary of the results.  You can search this website at any time.
WHAT ARE THE COSTS TO YOU IF YOU TAKE PART IN THIS STUDY? 
Page 8 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022


APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                 
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022
You will not be charged for any treatments or procedures that are part of this study.  If you 
usually pay co-payments for VA care and medications, you will still pay these co-payments for 
VA care and medications that are not part of this study.
IS THERE PAYMENT TO YOU IF YOU TAKE PART IN THIS STUDY?
 
     
You will receive $30 for the screening session, $30 for the script development session, and 
$150 for attending the test session. Thus, the total amount that you could receive during the 
study is $210.  This payment is for the time and effort associated with study assessments and 
procedures. You will not be paid if a test day is cancelled or if you test positive for alcohol or 
illicit drugs.  Payments will be given in the form of cash at the time of your participation.  If you 
participate in a virtual screening and are not eligible to come for the in-person visits you can 
choose to come in to pick up your cash payment, or be mailed a gift card or check.  If you 
choose to receive payment by gift card or check you may need to complete a tax form.
WHAT WILL HAPPEN IF YOU ARE INJURED BECAUSE OF YOUR BEING IN THE STUDY?
If you are injured as a result of taking part in this study, the VA will provide necessary medical 
treatment at no cost to you unless the injury is due to non-compliance by a study subject with 
study procedures. Emergency and ongoing medical treatment will be provided as needed.  
There are no plans to provide compensation for disability or other losses occurring over the long 
term or if an injury becomes apparent after your participation in the study has ended.  However, 
by agreeing to participate in this research study, you are not waiving or giving up any legal 
rights to seek compensation.  
If you have questions you can contact:
DURING THE DAY:
The VA Connecticut Research Coordinator at 203-937-3830, OR
Dr. Ralevski  at 203-932-5711 x4282 and 
AFTER HOURS:
The research pharmacist on call at 203-932-5711.                                  
Emergency and ongoing medical treatment will be provided as needed.  
WHO ELSE MAY YOU CONTACT IF YOU HAVE QUESTIONS?
Page 9 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022


APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                 
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022
If you have questions about your rights as a study subject, or you want to make sure this is a 
valid VA study, you may contact the Chairman of the Human Studies Subcommittee at 203-932-
5711 x3350.
If you have questions, complaints or concerns about the study or if you would like to obtain 
information or offer input you may call Dr. Ralevski at 203-932-5711 x4282.  
DO YOU HAVE TO TAKE PART IN THE STUDY?
Participation in this study is voluntary.  You may withdraw from this study at any time. Refusal to 
take part in the study, or withdrawing from the study, will involve no penalty or loss of benefits to 
which you are otherwise entitled. 
If you are a VA employee or student, the refusal to take part in the study will in no way influence 
your employment, ratings, subsequent recommendations, or academic progress as applicable.  
You may discontinue taking part at any time without any penalty or loss of benefits. 
RIGHT OF INVESTIGATOR TO TERMINATE YOUR PARTICIPATION 
You may be withdrawn from the study at the discretion of the investigators if you fail to follow 
instruction, do not meet entry criteria, or if the study physician feels it would be harmful for you 
to continue participating.  If you test positive for pregnancy, alcohol or illicit drugs you may be 
withdrawn from the study.  Blood pressure and heart rate are monitored frequently during the 
lab session and if they exceed the safety parameters you will be terminated from the study. The 
investigator may continue to review the data already collected for the study (prior to your 
withdrawal) but cannot collect further information, except from public records, such as survival 
data.  We do not anticipate any adverse effects on your health or welfare as a result of early 
termination.  
WILL YOU BE TOLD NEW INFORMATION ABOUT THIS STUDY? 
You and your physician will be informed of any important discoveries made during this study 
which may affect you, your condition, or your willingness to participate in this study.  If suicidal 
intent or other major clinical findings are detected during clinical or research assessments, the 
study physician will be notified immediately. You may be held against your will until you are 
judged to no longer be considered a threat to yourself or to others.
RE-CONTACT
We will re-contact after the original project is complete if you are interested in the findings of this 
study.
Page 10 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022


APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022
RESEARCH CONSENT  FORM
VA Connecticut Healthcare System
Subject Name:          ___________________                                                                  Date:  _________
Title of Study: _Effects of allopregnanolone on stress-induced craving                 
Principal Investigator:  __Elizabeth Ralevski, PhD_________ Version Date: _03/09/2022
If you are willing to be contacted for future research by our group, please specify:
YES _________         No____________
AGREEMENT TO PARTICIPATE IN THE RESEARCH STUDY
The research staff has explained  the research study to you.  You have been told of the risks or 
discomforts and possible benefits of the study. You have been told of other choices of treatment 
available to you.  You have been given the chance to ask questions and obtain answers.  
By signing this document below, you voluntarily consent to participate in this study You also 
confirm that you have read this consent document, or it has been read to you.  You will receive 
a copy of this consent document after you sign it.  
I agree to participate in this research study as it has been explained in this document.
_________________________
Subject’s Name
____________________________
Subject’s Signature
___________
Date
_________________________
Person Obtaining Consent
____________________________
Person Obtaining: Signature
___________
Date
Page 11 of 11
 
VA Connecticut Healthcare System Human Studies
Subcommittee
Effective Date:
March 17, 2022
APPROVED BY THE YALE UNIVERSITY IRB 4/6/2022