Trial Outcomes & Findings for Immune Response and General Immune Health in Subjects Infected With Herpes Simplex Virus Type 1 (HSV-1) (NCT NCT03661541)
NCT ID: NCT03661541
Last Updated: 2021-11-03
Results Overview
RNA gene expression of interleukin-5 (IL5) and interferon gamma (IFNG) genes in PBMCs exposed in vitro to 4 conditions: no stimulus, HSV-1 virus, HSV-1-infected cell extracts, and Candida albicans extract. Measure is qRT-PCR crossing threshold (Ct), where a lower number is higher gene expression. A lower Ct of 1 is approximately a 2-fold higher expression level of the gene.
Recruitment status
COMPLETED
Study phase
PHASE1
Target enrollment
36 participants
Primary outcome timeframe
Baseline and 8 weeks after baseline (Day 1) +/- 7 days for subjects with less than 6 herpres labialis outbreaks in 12 months, relative to baseline for subjects with 6 or greater herpes labialis outbreaks
Results posted on
2021-11-03
Participant Flow
Participant milestones
| Measure |
6 or More Herpes Labialis Outbreaks
Subjects will be recruited in three groups, all of whom are infected with HSV-1, as shown by having IgG against HSV-1:
Group A: 12 subjects with 6 or more herpes labialis outbreaks in the past 12 months.
Subjects in group A will receive 2% Squaric Acid Dibutyl Ester (SADBE) dose on the arm after their initial blood samples are obtained. Group A subjects will have blood collected and tests repeated 2 and 8 weeks later.
Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges.
Squaric Acid Dibutyl Ester: 2% squaric acid dibutyl ester (SADBE) (Supplied by Squarex) is topically applied to the inner aspect of the upper arm of the subject and covered with Tegaderm. Subject is advised to wash it off after 3 hours.
|
1 to 2 Herpes Labialis Outbreaks
Subjects will be recruited in three groups, all of whom are infected with HSV-1, as shown by having IgG against HSV-1:
Group B: 12 subjects with 1 to 2 outbreaks in the past 12 months. Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges. Invite these back for a further blood draw on Day 0.
|
Zero Herpes Labialis Outbreaks
Subjects will be recruited in three groups, all of whom are infected with HSV-1, as shown by having IgG against HSV-1:
Group C: 12 subjects with zero outbreaks in the past 12 months. Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges. Invite these back for a further blood draw on Day 0.
|
|---|---|---|---|
|
Overall Study
STARTED
|
12
|
12
|
12
|
|
Overall Study
COMPLETED
|
10
|
10
|
10
|
|
Overall Study
NOT COMPLETED
|
2
|
2
|
2
|
Reasons for withdrawal
Withdrawal data not reported
Baseline Characteristics
Immune Response and General Immune Health in Subjects Infected With Herpes Simplex Virus Type 1 (HSV-1)
Baseline characteristics by cohort
| Measure |
6 or More Herpes Labialis Outbreaks
n=12 Participants
Subjects will be recruited in three groups, all of whom are infected with HSV-1, as shown by having IgG against HSV-1:
Group A: 12 subjects with 6 or more herpes labialis outbreaks in the past 12 months.
Subjects in group A will receive 2% Squaric Acid Dibutyl Ester (SADBE) dose on the arm after their initial blood samples are obtained. Group A subjects will have blood collected and tests repeated 2 and 8 weeks later.
Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges.
Squaric Acid Dibutyl Ester: 2% squaric acid dibutyl ester (SADBE) (Supplied by Squarex) is topically applied to the inner aspect of the upper arm of the subject and covered with Tegaderm. Subject is advised to wash it off after 3 hours.
|
1 or 2 Herpes Labialis Outbreaks
n=12 Participants
Subjects will be recruited in three groups, all of whom are infected with HSV-1, as shown by having IgG against HSV-1:
Group B: 12 subjects with 1 to 2 outbreaks in the past 12 months. Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges. Invite these back for a further blood draw on Day 0.
|
Zero Herpes Labialis Outbreaks
n=12 Participants
Subjects will be recruited in three groups, all of whom are infected with HSV-1, as shown by having IgG against HSV-1:
Group C: 12 subjects with zero outbreaks in the past 12 months. Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges. Invite these back for a further blood draw on Day 0.
|
Total
n=36 Participants
Total of all reporting groups
|
|---|---|---|---|---|
|
Age, Categorical
<=18 years
|
0 Participants
n=93 Participants
|
0 Participants
n=4 Participants
|
0 Participants
n=27 Participants
|
0 Participants
n=483 Participants
|
|
Age, Categorical
Between 18 and 65 years
|
12 Participants
n=93 Participants
|
12 Participants
n=4 Participants
|
12 Participants
n=27 Participants
|
36 Participants
n=483 Participants
|
|
Age, Categorical
>=65 years
|
0 Participants
n=93 Participants
|
0 Participants
n=4 Participants
|
0 Participants
n=27 Participants
|
0 Participants
n=483 Participants
|
|
Age, Continuous
|
51.71 years
n=93 Participants
|
51.71 years
n=4 Participants
|
54.30 years
n=27 Participants
|
52.57 years
n=483 Participants
|
|
Sex: Female, Male
Female
|
9 Participants
n=93 Participants
|
9 Participants
n=4 Participants
|
8 Participants
n=27 Participants
|
26 Participants
n=483 Participants
|
|
Sex: Female, Male
Male
|
3 Participants
n=93 Participants
|
3 Participants
n=4 Participants
|
4 Participants
n=27 Participants
|
10 Participants
n=483 Participants
|
|
Race (NIH/OMB)
American Indian or Alaska Native
|
0 Participants
n=93 Participants
|
1 Participants
n=4 Participants
|
0 Participants
n=27 Participants
|
1 Participants
n=483 Participants
|
|
Race (NIH/OMB)
Asian
|
0 Participants
n=93 Participants
|
1 Participants
n=4 Participants
|
0 Participants
n=27 Participants
|
1 Participants
n=483 Participants
|
|
Race (NIH/OMB)
Native Hawaiian or Other Pacific Islander
|
0 Participants
n=93 Participants
|
0 Participants
n=4 Participants
|
0 Participants
n=27 Participants
|
0 Participants
n=483 Participants
|
|
Race (NIH/OMB)
Black or African American
|
1 Participants
n=93 Participants
|
1 Participants
n=4 Participants
|
3 Participants
n=27 Participants
|
5 Participants
n=483 Participants
|
|
Race (NIH/OMB)
White
|
11 Participants
n=93 Participants
|
9 Participants
n=4 Participants
|
9 Participants
n=27 Participants
|
29 Participants
n=483 Participants
|
|
Race (NIH/OMB)
More than one race
|
0 Participants
n=93 Participants
|
0 Participants
n=4 Participants
|
0 Participants
n=27 Participants
|
0 Participants
n=483 Participants
|
|
Race (NIH/OMB)
Unknown or Not Reported
|
0 Participants
n=93 Participants
|
0 Participants
n=4 Participants
|
0 Participants
n=27 Participants
|
0 Participants
n=483 Participants
|
|
Region of Enrollment
United States
|
12 participants
n=93 Participants
|
12 participants
n=4 Participants
|
12 participants
n=27 Participants
|
36 participants
n=483 Participants
|
PRIMARY outcome
Timeframe: Baseline and 8 weeks after baseline (Day 1) +/- 7 days for subjects with less than 6 herpres labialis outbreaks in 12 months, relative to baseline for subjects with 6 or greater herpes labialis outbreaksPopulation: Absolute gene expression for Group A on Day 57, Group B on day 1 and Group C on day 1 compared to Group A on Day 1.
RNA gene expression of interleukin-5 (IL5) and interferon gamma (IFNG) genes in PBMCs exposed in vitro to 4 conditions: no stimulus, HSV-1 virus, HSV-1-infected cell extracts, and Candida albicans extract. Measure is qRT-PCR crossing threshold (Ct), where a lower number is higher gene expression. A lower Ct of 1 is approximately a 2-fold higher expression level of the gene.
Outcome measures
| Measure |
Group A: 6 or More Herpes Labialis Outbreaks, Day 1
n=12 Participants
Group A: 12 subjects positive for Ab against HSV-1 and self-reporting 6 or more herpes labialis outbreaks in the past 12 months.
Subjects in group A will receive 2% Squaric Acid Dibutyl Ester (SADBE) dose on the arm after their initial blood samples on day 1 are obtained. Group A subjects had blood collected and tests repeated 2 and 8 weeks later.
Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges.
Squaric Acid Dibutyl Ester: 2% squaric acid dibutyl ester (SADBE) (Supplied by Squarex) is topically applied to the inner aspect of the upper arm of the subject and covered with Tegaderm. Subject is advised to wash it off after 3 hours.
|
Group B: 1 or 2 Herpes Labialis Outbreaks in Prior 12 Months, Day 1
n=12 Participants
Group B: Subjects positive for Ab against HSV-1 and self-reporting 1 or 2 herpes labialis outbreaks in the prior 12 months.
Group B: 12 subjects with 1 to 2 outbreaks in the past 12 months. Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges. Results from blood draw on day 1.
|
Group C: Zero Herpes Labialis Outbreaks in Prior 12 Months, Day 1
n=12 Participants
Group C: 12 subjects positive for Ab against HSV-1 nad self-reporting zero herpes labialis outbreaks in the past 12 months. Blood draw is on day 1. Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges.
|
Group A, Day 57. Subjects With 6 or More Outbreaks, 8 Weeks After Single Dose of SADBE.
n=12 Participants
Group A, day 57. Subjects positive for Ab against HSV-1, self-reporting 6 or more herpes labialis outbreaks in the prior 12 months. Blood was collected on day 57 after a single topical dose of SADBE applied to the upper arm on day 1.
|
|---|---|---|---|---|
|
Ratio of Absolute Gene Expression of IFNG and IL5 in PBMCs Between 8 Weeks and Baseline
IL5 in negative control
|
10.073 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 0.544
|
11.380 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.233
|
10.711 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.274
|
11.431 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.143
|
|
Ratio of Absolute Gene Expression of IFNG and IL5 in PBMCs Between 8 Weeks and Baseline
IL5 with HSV-1-infected cell extract stimulus
|
9.624 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 0.666
|
10.983 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 0.971
|
10.986 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.268
|
10.627 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 0.810
|
|
Ratio of Absolute Gene Expression of IFNG and IL5 in PBMCs Between 8 Weeks and Baseline
IL5 with HSV-1 virus stimulus
|
9.747 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 0.697
|
10.736 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 0.880
|
10.414 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.217
|
10.595 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 0.957
|
|
Ratio of Absolute Gene Expression of IFNG and IL5 in PBMCs Between 8 Weeks and Baseline
IL5 with Candida stimulus
|
10.680 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 0.941
|
10.806 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.093
|
11.051 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.332
|
10.536 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.215
|
|
Ratio of Absolute Gene Expression of IFNG and IL5 in PBMCs Between 8 Weeks and Baseline
IFNG in negative control
|
10.033 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 0.820
|
10.210 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.320
|
9.729 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.902
|
9.718 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.164
|
|
Ratio of Absolute Gene Expression of IFNG and IL5 in PBMCs Between 8 Weeks and Baseline
IFNG in HSV-1-infected cell extracts stimulus
|
6.040 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.681
|
3.898 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.900
|
4.992 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 2.418
|
3.522 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.600
|
|
Ratio of Absolute Gene Expression of IFNG and IL5 in PBMCs Between 8 Weeks and Baseline
IFNG in HSV-1 stimulus
|
7.384 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.670
|
5.554 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 2.650
|
5.789 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.923
|
3.923 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.548
|
|
Ratio of Absolute Gene Expression of IFNG and IL5 in PBMCs Between 8 Weeks and Baseline
IFNG in Candida stimulus
|
8.421 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 2.053
|
6.167 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 2.980
|
7.336 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 1.957
|
5.327 q-RT-PCR crossing threshold cycle (Ct)
Standard Deviation 2.696
|
PRIMARY outcome
Timeframe: BaselinePBMC proliferation in response to 3 stimuli: HSV-1 virus, HSV-1-infected cell extracts, and Candida albicans extract.
Outcome measures
| Measure |
Group A: 6 or More Herpes Labialis Outbreaks, Day 1
n=12 Participants
Group A: 12 subjects positive for Ab against HSV-1 and self-reporting 6 or more herpes labialis outbreaks in the past 12 months.
Subjects in group A will receive 2% Squaric Acid Dibutyl Ester (SADBE) dose on the arm after their initial blood samples on day 1 are obtained. Group A subjects had blood collected and tests repeated 2 and 8 weeks later.
Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges.
Squaric Acid Dibutyl Ester: 2% squaric acid dibutyl ester (SADBE) (Supplied by Squarex) is topically applied to the inner aspect of the upper arm of the subject and covered with Tegaderm. Subject is advised to wash it off after 3 hours.
|
Group B: 1 or 2 Herpes Labialis Outbreaks in Prior 12 Months, Day 1
n=12 Participants
Group B: Subjects positive for Ab against HSV-1 and self-reporting 1 or 2 herpes labialis outbreaks in the prior 12 months.
Group B: 12 subjects with 1 to 2 outbreaks in the past 12 months. Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges. Results from blood draw on day 1.
|
Group C: Zero Herpes Labialis Outbreaks in Prior 12 Months, Day 1
n=12 Participants
Group C: 12 subjects positive for Ab against HSV-1 nad self-reporting zero herpes labialis outbreaks in the past 12 months. Blood draw is on day 1. Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges.
|
Group A, Day 57. Subjects With 6 or More Outbreaks, 8 Weeks After Single Dose of SADBE.
Group A, day 57. Subjects positive for Ab against HSV-1, self-reporting 6 or more herpes labialis outbreaks in the prior 12 months. Blood was collected on day 57 after a single topical dose of SADBE applied to the upper arm on day 1.
|
|---|---|---|---|---|
|
Immune Monitoring - PBMC Proliferation
HSV-1-infected cell extracts
|
16.73 percent proliferation
Standard Error 3.94
|
19.26 percent proliferation
Standard Error 5.57
|
20.19 percent proliferation
Standard Error 4.47
|
—
|
|
Immune Monitoring - PBMC Proliferation
HSV-1 virus
|
8.26 percent proliferation
Standard Error 1.98
|
11.71 percent proliferation
Standard Error 2.98
|
12.70 percent proliferation
Standard Error 3.05
|
—
|
|
Immune Monitoring - PBMC Proliferation
Candida extract
|
4.47 percent proliferation
Standard Error 1.53
|
4.85 percent proliferation
Standard Error 1.51
|
7.83 percent proliferation
Standard Error 2.19
|
—
|
|
Immune Monitoring - PBMC Proliferation
Normalized all three stimuli
|
68.33 percent proliferation
Standard Error 10.42
|
83.21 percent proliferation
Standard Error 13.53
|
99.98 percent proliferation
Standard Error 13.92
|
—
|
PRIMARY outcome
Timeframe: Day 1 and 8 weeks from Day 1 (Initial visit) +/- 7 days for subjects with 6 or more herpes labalis outbreaksPopulation: Only subjects with 6 or more herpes labialis outbreaks were treated, therefore only subjects with 6 or more outbreaks are included in this analysis. This set of data compares them at Day 1, before treatment, and analyzed at 8 weeks from Day 1 treatment. Data were not collected at 8 weeks on the subjects with 1 or 2 outbreaks or with zero outbreaks. Data at day 1 on those groups and the subjects with 6 or more outbreaks is reported in Outcome Measure 2.
PBMC proliferation in response to 3 stimuli: HSV-1 virus, HSV-1-infected cell extracts, and Candida albicans extract.
Outcome measures
| Measure |
Group A: 6 or More Herpes Labialis Outbreaks, Day 1
n=12 Participants
Group A: 12 subjects positive for Ab against HSV-1 and self-reporting 6 or more herpes labialis outbreaks in the past 12 months.
Subjects in group A will receive 2% Squaric Acid Dibutyl Ester (SADBE) dose on the arm after their initial blood samples on day 1 are obtained. Group A subjects had blood collected and tests repeated 2 and 8 weeks later.
Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges.
Squaric Acid Dibutyl Ester: 2% squaric acid dibutyl ester (SADBE) (Supplied by Squarex) is topically applied to the inner aspect of the upper arm of the subject and covered with Tegaderm. Subject is advised to wash it off after 3 hours.
|
Group B: 1 or 2 Herpes Labialis Outbreaks in Prior 12 Months, Day 1
n=12 Participants
Group B: Subjects positive for Ab against HSV-1 and self-reporting 1 or 2 herpes labialis outbreaks in the prior 12 months.
Group B: 12 subjects with 1 to 2 outbreaks in the past 12 months. Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges. Results from blood draw on day 1.
|
Group C: Zero Herpes Labialis Outbreaks in Prior 12 Months, Day 1
Group C: 12 subjects positive for Ab against HSV-1 nad self-reporting zero herpes labialis outbreaks in the past 12 months. Blood draw is on day 1. Subjects in Groups B and C will be matched to the subjects in Group A so that demographic characteristics (i.e., gender distribution, age and weight) are within broadly similar ranges.
|
Group A, Day 57. Subjects With 6 or More Outbreaks, 8 Weeks After Single Dose of SADBE.
Group A, day 57. Subjects positive for Ab against HSV-1, self-reporting 6 or more herpes labialis outbreaks in the prior 12 months. Blood was collected on day 57 after a single topical dose of SADBE applied to the upper arm on day 1.
|
|---|---|---|---|---|
|
Immune Monitoring - PBMC Proliferation
HSV-1-infected cell extracts
|
16.96 percent proliferation
Standard Error 2.62
|
16.73 percent proliferation
Standard Error 3.94
|
—
|
—
|
|
Immune Monitoring - PBMC Proliferation
HSV-1 virus
|
13.37 percent proliferation
Standard Error 2.38
|
8.26 percent proliferation
Standard Error 1.98
|
—
|
—
|
|
Immune Monitoring - PBMC Proliferation
Candida extract
|
7.35 percent proliferation
Standard Error 2.42
|
4.47 percent proliferation
Standard Error 1.53
|
—
|
—
|
|
Immune Monitoring - PBMC Proliferation
Normalized all three stimuli
|
99.97 percent proliferation
Standard Error 13.11
|
73.75 percent proliferation
Standard Error 11.56
|
—
|
—
|
Adverse Events
6 or More Herpes Labialis Outbreaks
Serious events: 0 serious events
Other events: 3 other events
Deaths: 0 deaths
Serious adverse events
Adverse event data not reported
Other adverse events
| Measure |
6 or More Herpes Labialis Outbreaks
n=12 participants at risk
Subjects will be recruited in three groups, all of whom are infected with HSV-1, as shown by having IgG against HSV-1:
Group A: 12 subjects with 6 or more herpes labialis outbreaks in the past 12 months.
Subjects in group A will receive 2% Squaric Acid Dibutyl Ester (SADBE) dose on the arm after their initial blood samples are obtained. Group A subjects will have blood collected and tests repeated 2 and 8 weeks later.
Group B and C subjects (having 0 or 1 to 2 outbreaks in the prior 12 months) were not treated, and therefore not at risk, and adverse event data was not collected on them.
Squaric Acid Dibutyl Ester: 2% squaric acid dibutyl ester (SADBE) (Supplied by Squarex) is topically applied to the inner aspect of the upper arm of the subject and covered with Tegaderm. Subject is advised to wash it off after 3 hours.
Only subjects with 6 or more herpes labialis outbreaks (Group A) were treated and therefore, only this group is included in the adverse event reporting. The patients in the zero outbreaks group and 1 to 2 outbreaks group were not treated with either drug or placebo, and therefore not at risk of adverse events, and they were not followed for adverse events and no adverse event data was collected for those groups. So, again, only subjects in Group A (6 or more outbreaks in the prior 12 months) were treated and thus at risk for adverse events, and adverse event data was collected only for that group.
|
|---|---|
|
Skin and subcutaneous tissue disorders
Local reaction
|
16.7%
2/12 • Number of events 2 • Adverse events were collected over 8 weeks
Only subjects with 6 or more herpes labialis outbreaks (Group A) were treated and therefore, only this group is included in the adverse event reporting. Subjects in the other two groups were not treated with either drug or placebo, and therefore not at risk of adverse events, and they were not followed for adverse events and no adverse event data was collected for those groups.
|
|
Gastrointestinal disorders
Nausea
|
8.3%
1/12 • Number of events 1 • Adverse events were collected over 8 weeks
Only subjects with 6 or more herpes labialis outbreaks (Group A) were treated and therefore, only this group is included in the adverse event reporting. Subjects in the other two groups were not treated with either drug or placebo, and therefore not at risk of adverse events, and they were not followed for adverse events and no adverse event data was collected for those groups.
|
Additional Information
Results disclosure agreements
- Principal investigator is a sponsor employee
- Publication restrictions are in place