Effectiveness Of Ceftriaxone and

N-acetylcysteine on Nicotine Withdrawal and

Nicotine-induced Reinstatement of Preference
in Sprague–Dawley Rats
 Declaration of Conflict of Interest
 We declare that we do not have any conflict of interest in this
undertaking.
 This research study is self-financed and is not intended to advertise
any pharmaceutical company.
 This study was done in partial fulfillment of the requirements in our
Pharmacology Course.
Introduction
 Introduction

 Smoking is one of the major preventable

causes of morbidity and mortality
worldwide.
 Gap in Knowledge

• Despite some policies and pharmacologic

interventions addressing the negative
effects of tobacco addiction and nicotine
abuse, cigarette use and relapse rates are
still alarming at present.
 Ceftriaxone

• attenuate the reinstatement of nicotine

• enhance nicotine-induced anti-nociception
• attenuate the development of chronic nicotine
tolerance in mice.

Sondheimer I, Knackstedt L. Ceftriaxone prevents the induction of cocaine sensitization and produces enduring attenuation of
cue- and cocaine-primed reinstatement of cocaine-seeking. Behav Brain Res. 2011 November; 225(1).
 N-Acetylcysteine

• attenuate perceived reward from smoking

following an adlib-smoking period.

Schmaal L, Berk L, Hulstijn K, Cousijn J, Wiers R, van den Brink W. Efficacy of N-Acetylcysteine in the Treatment of
Nicotine Dependence: A Double-Blind Placebo-Controlled Pilot Study. Eur Addict Res. 2011;17(4):211-216.
 Objective
• to compare the effectiveness of Ceftriaxone and
N-acetylcysteine on nicotine withdrawal and
nicotine induced reinstated preference in
Sprague–Dawley rats.
Methodology
 Methodology

• Experimental Research
• Observer-Blind
• Research Animal: Sprague-Dawley Rats
Flowchart of the Experiment Proper
Group Selection and Acclimatization
Conditioned Place Preference (CPP) Test

Nicotine Addiction Phase

Nicotine-primed CPP Test Tail Immersion Test Hot Plate Test

Treatment and Withdrawal Phase

Elevated Maze Test Tail Immersion Test Hot Plate Test

Relapse Phase
Nicotine-primed CPP Test
Flowchart of the Experiment Proper
Group Selection and Acclimatization
Conditioned Place Preference (CPP) Test

Nicotine Addiction Phase

Nicotine-primed CPP Test Tail Immersion Test Hot Plate Test

Treatment and Withdrawal Phase

Elevated Maze Test Tail Immersion Test Hot Plate Test

Relapse Phase
Nicotine-primed CPP Test
Group Selection and Acclimatization
Conditioned Place Preference Test
Flowchart of the Experiment Proper
Group Selection and Acclimatization
Conditioned Place Preference (CPP) Test

Nicotine Addiction Phase

Nicotine-primed CPP Test Tail Immersion Test Hot Plate Test

Treatment and Withdrawal Phase

Elevated Maze Test Tail Immersion Test Hot Plate Test

Relapse Phase
Nicotine-primed CPP Test
 Nicotine Addiction Phase

• Nicotine Injections (SQ):

• Days 1-5: ¼ Dose
• Days 6-10: ½ Dose
• Days 11-15: Full Dose (0.9mg/kg)

• Saline Injections: 1cc/day for the Saline Group

 Nicotine Addiction Phase
Nicotine-primed CPP Test Tail Immersion Test Hot Plate Test
Flowchart of the Experiment Proper
Group Selection and Acclimatization
Conditioned Place Preference (CPP) Test

Nicotine Addiction Phase

Nicotine-primed CPP Test Tail Immersion Test Hot Plate Test

Treatment and Withdrawal Phase

Elevated Maze Test Tail Immersion Test Hot Plate Test

Relapse Phase
Nicotine-primed CPP Test
 Withdrawal & Treatment Phase
• Treatments Given (SQ):
• Varenicline- 0.03 mg/kg
• Ceftriaxone- 90 mg/kg
• N-acetylcysteine- 90 mg/kg

• Saline Injections: 90mg/kg for the Saline Group

 Treatment and Withdrawal Phase
Elevated Maze Test Tail Immersion Test Hot Plate Test
Flowchart of the Experiment Proper
Group Selection and Acclimatization
Conditioned Place Preference (CPP) Test

Nicotine Addiction Phase

Nicotine-primed CPP Test Tail Immersion Test Hot Plate Test

Treatment and Withdrawal Phase

Elevated Maze Test Tail Immersion Test Hot Plate Test

Relapse Phase
Nicotine-primed CPP Test
Results and Discussion
 Addiction Phase

Table 1. Mean time of the rats in the nicotine/intervention box (N=20)

Group n Average Time (seconds)

Saline 5 148*
Nicotine 15 238*

*p = 0.001
 Addiction Phase

Table 2. Average reaction times for the Tail Immersion and Hot Plate tests
(N=20)
Tail Immersion Test Hot Plate Test
Group n
(Seconds) (Seconds)
Saline 5 10.68 22.21
Nicotine 15 10.94 23.71
p = 0.815 p = 0.455
 Treatment and Withdrawal Phase
Table 3. Average behaviors on the elevated plus maze (N=20)
Time until first Number of
Open Arm Time
Group n response Somatic Signs
(Seconds)
(Seconds) (Average)
Saline 5 43.27 77.01 17
Varenicline 5 28.36 60.98 22
Ceftriaxone 5 30.61 55.70 28
N-acetylcysteine 5 24.59 66.04 32

p = 0.155 p = 0.705 p = 0.189

 Treatment and Withdrawal Phase
Table 4. Average reaction times for the Tail Immersion and Hot Plate tests
(N=20)
Tail Immersion Test Hot Plate Test
Group n
(Seconds) (Seconds)
Saline 5 10.67 17.46
Varenicline 5 8.74 25.93
Ceftriaxone 5 8.05 18.08
N-acetylcysteine 5 8.30 25.96
p = 0.099 p = 0.285
 Relapse Phase

Table 5. Mean time of the rats in the nicotine/intervention box post-

treatment (N=20)
Group n Average Time (seconds)
Saline 5 310.40
Varenicline 5 204.40
Ceftriaxone 5 167.40
N-acetylcysteine 5 264.20
p = 0.545
 Discussion

• Results corroborated with the findings made by:

 Aljaji et. al. in 2013 regarding Ceftriaxone
Frogeliger, Sohmaal, and Murray et. al. regarding N-
acetylcysteine.
 Discussion: Ceftriaxone
• “xCT and GLT-1 are down-regulated in the nucleus accumbens by
nicotine the present data supports the potential relevance in the
development and/or expression of dependence.”

Alajaji M, Bowers M, Knackstedt L, Damaj M. Effects of the beta-lactam antibiotic Ceftriaxone on

nicotine withdrawal and nicotine-induced reinstatement of preference in rats. Psychopharmacology.
2013;228(3):419-426.
 Discussion: N-acetylcysteine
• “The NAC group reported less nicotine-withdrawal symptoms and
maintained abstinence”

Froeliger B et al. The effects of N-acetylcysteine on frontostriatal resting-state functional connectivity,

withdrawal symptoms and smoking abstinence: A double-blind, placebo-controlled fMRI pilot study. Drug
ALcohol Dependence. 2015 November
 Discussion: N-acetylcysteine
• “…its capacity to inhibit dopamine release and reduce synaptic
release of glutamate by increasing extracellular levels of glutamate
therefore reducing the reward behavior associated with nicotine
addiction.”

Murray J, Lacoste J, Belin D. N-Acetylcysteine as a Treatment for Addiction [Internet]. InTech.

2012 [cited 22 May 2016]. Available from: http://cdn.intechopen.com/pdfs/40326/InTech-
N_acetylcysteine_as_a_treatment_for_addiction.pdf
Conclusion
 Conclusion

• the results showed that both ceftriaxone and N-

acetylcysteine were able to eliminate the expression
of somatic and nociceptic withdrawal signs in nicotine-
dependent mice.
 Conclusion

• the effectiveness of Ceftriaxone and N-acetylcysteine

was comparable based on nicotine withdrawal using
varenicline and nicotine-induced reinstated
preference in Sprague–Dawley rats
 Conclusion

• This signifies that ceftriaxone and N-acetylcysteine

may be used as alternative drugs to varenicline in
nicotine cessation therapy.
Recommendations
 Recommendation

 Other potential drugs that could elicit the same

effects are encouraged to be studied alongside
the drugs used in the study to further improve
their potential as management for nicotine
withdrawal.
 References
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Dependence: A Double-Blind Placebo-Controlled Pilot Study. Eur Addict Res. 2011;17(4):211-216.
3. Alajaji M, Bowers M, Knackstedt L, Damaj M. Effects of the beta-lactam antibiotic Ceftriaxone on nicotine withdrawal and
nicotine-induced reinstatement of preference in rats. Psychopharmacology. 2013;228(3):419-426.
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