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A Monoclonal Anti–Calcitonin Gene-Related
Peptide Antibody Decreases Stress-Induced
Colonic Hypersensitivity
Ehsan Noor-Mohammadi et al.
Department of Physiology
University of Oklahoma Health Science Center
Oklahoma City, Oklahoma
Teva Pharmaceutical Industries, Ltd., Redwood City, California
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Year 2021, Volume 379, Issue 3 [page 270-279]
Irritable Bowel Syndrome
2
➢ Brain-gut disorder
➢ Clinical symptoms:
▪ Abdominal pain and
discomfort
▪ Bloating
▪ Abnormal bowel habits
Cause
Calcitonin Gene Related Peptide
3
➢37-amino-acid member of the calcitonin family of peptides
➢Produced: peripheral and central neurons
➢Potent vasodilator
➢Function: Transmission of nociception
GI: Motility, secretion, and neuroprotection
Anti-CGRP Antibody
4
➢Inflammatory models : CGRP receptor antagonist - visceral
hypersensitivity.
➢Anti-CGRP divalent antigen-
binding fragment [F(ab’)2]
antibody
Material & Methods
5
➢ Drug: Fremanezumab (Monoclonal Anti-CGRP antibody)
Divalent antigen-binding fragment [F(ab‘)2] antibody
30mg/kg Intraperitoneal
➢ Animal
Male: Fischer 344 Female: Long Evan
Sprague Dawley rat
Material & Methods
6
➢ Chronic Adult Stress Induced by Water Avoidance Stress
➢ Odor-Attachment Learning Model of ELS: Peppermint oil
➢ Colonic Sensitivity: Distender series IIR barostat
➢ pERK Immunohistochemistry
Material & Methods
7
Rodent Olfactometer Distender series IIR barostat
Y Maze
Experimental Design
8
Water Avoidance stress + Vehicle
Water Avoidance stress + Isotype F(ab’)2 Control
Water Avoidance stress + Anti-CGRP F(ab’)2
SHAM + Vehicle
SHAM + Isotype F(ab’)2 Control
SHAM + Anti-CGRP F(ab’)2
Experimental Design
9
Unpredictable + Vehicle
Unpredictable + Isotype F(ab’)2 Control
Unpredictable + Anti-CGRP F(ab’)2
Odor only + Vehicle
Odor only + Isotype F(ab’)2 Control
Odor only + Anti-CGRP F(ab’)2
Results
10
1. Pharmacokinetics of Anti-CGRP F(ab’)2
Results
11
2. Anti-CGRP F(ab’)2 Inhibits Colonic Hypersensitivity Induced by WAS
without an Effect on Colonic Compliance
Two way ANOVA test followed by Bonferroni post test.
Results
12
3. Anti-CGRP F(ab’)2 Inhibits Thoracolumbar Spinal Cord Signaling Induced by
CRD in Rats Previously Exposed to WAS
0
1
2
3
4
5
Vehicle Isotype F(ab')2 Anti-CGRP
F(ab')2
T10-12
SHAM WAS
Average
Number
of
pERK-IR
cells/section
***
***
####
***P < 0.001 and ****P < 0.0001 compared with WAS 1 VEH; #### P < 0.0001 compared WAS 1 isotype F(ab’)2 control.
Results
13
3. Anti-CGRP F(ab’)2 Inhibits Thoracolumbar Spinal Cord Signaling Induced by
CRD in Rats Previously Exposed to WAS
Average
Number
of
pERK-IR
cells/section
0
1
2
3
4
5
Vehicle Isotype
F(ab')2
Anti-CGRP
F(ab')2
T13-L1
SHAM WAS
####
***P < 0.001 and ****P < 0.0001 compared with WAS 1 VEH; #### P < 0.0001 compared WAS 1 isotype F(ab’)2 control.
****
****
Results
14
4. Rodent Model of ELS: Behavioral Activation, Odor Preference, and
Weaning Weights
Results
15
4. Rodent Model of ELS: Behavioral Activation, Odor Preference, and
Weaning Weights
0
1
2
3
4
5
Odor Only Unpredictable
Odor Preference
Odor
Choices
0
10
20
30
40
50
Odor only Unpredictable
Weaning Weights
Weight
(gm)
Results
16
5. Anti-CGRP F(ab’)2 Inhibits Colonic Hypersensitivity in Adult Animals after
Neonatal Unpredictable ELS without an Effect on Colonic Compliance
Results
17
6. Anti-CGRP F(ab’)2 Inhibits Thoracolumbar Spinal Cord Signaling Induced
by CRD in Rats Previously Exposed to Neonatal Unpredictable ELS
0
2
4
6
8
Vehicle Isotype
F(ab')2
Anti-CGRP
F(ab')2
T10-12
Odor only Unpredictable
Average
Number
of
pERK-IR
cells/section
****
****
####
***P < 0.001 and ****P < 0.0001 compared with WAS 1 VEH; #### P < 0.0001 compared WAS 1 isotype F(ab’)2 control.
Results
18
6. Anti-CGRP F(ab’)2 Inhibits Thoracolumbar Spinal Cord Signaling Induced
by CRD in Rats Previously Exposed to Neonatal Unpredictable ELS
0
1
2
3
4
5
Vehicle Isotype
F(ab')2
Anti-CGRP
F(ab')2
T13-L1
Odor only Unpredictable
Average
Number
of
pERK-IR
cells/section
****
***
####
***P < 0.001 and ****P < 0.0001 compared with WAS 1 VEH; #### P < 0.0001 compared WAS 1 isotype F(ab’)2 control.
Discussion
19
To evaluate whether antibody mediated peripheral blockade
of CGRP prevented stress induced visceral pain in adulthood.
Repeated WAS
Unpredictable ELS
Peripheral CGRP
Colonic
hypersensitivity
Neuronal
activation
Dorsal horn of the
thoracolumbar spinal
cord
CGRP signalling
ELS-induced visceral
hypersensitivity
Conclusion
20
IBS : Multifaceted disorder Stress
early life
adulthood
F(ab’)2 fragment
antibody
Targeting CGRP inhibits
stress-induced colonic
hypersensitivity
Stimulus-evoked spinal
ERK1/2 phosphorylation
Regulation
of
persistent
visceral
pain
Discussion
21
Migraine
22
Discussion

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Dr Jeenal Mistry_Journal Club 2_12th March 2022.pdf

  • 1. A Monoclonal Anti–Calcitonin Gene-Related Peptide Antibody Decreases Stress-Induced Colonic Hypersensitivity Ehsan Noor-Mohammadi et al. Department of Physiology University of Oklahoma Health Science Center Oklahoma City, Oklahoma Teva Pharmaceutical Industries, Ltd., Redwood City, California THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS Year 2021, Volume 379, Issue 3 [page 270-279]
  • 2. Irritable Bowel Syndrome 2 ➢ Brain-gut disorder ➢ Clinical symptoms: ▪ Abdominal pain and discomfort ▪ Bloating ▪ Abnormal bowel habits Cause
  • 3. Calcitonin Gene Related Peptide 3 ➢37-amino-acid member of the calcitonin family of peptides ➢Produced: peripheral and central neurons ➢Potent vasodilator ➢Function: Transmission of nociception GI: Motility, secretion, and neuroprotection
  • 4. Anti-CGRP Antibody 4 ➢Inflammatory models : CGRP receptor antagonist - visceral hypersensitivity. ➢Anti-CGRP divalent antigen- binding fragment [F(ab’)2] antibody
  • 5. Material & Methods 5 ➢ Drug: Fremanezumab (Monoclonal Anti-CGRP antibody) Divalent antigen-binding fragment [F(ab‘)2] antibody 30mg/kg Intraperitoneal ➢ Animal Male: Fischer 344 Female: Long Evan Sprague Dawley rat
  • 6. Material & Methods 6 ➢ Chronic Adult Stress Induced by Water Avoidance Stress ➢ Odor-Attachment Learning Model of ELS: Peppermint oil ➢ Colonic Sensitivity: Distender series IIR barostat ➢ pERK Immunohistochemistry
  • 7. Material & Methods 7 Rodent Olfactometer Distender series IIR barostat Y Maze
  • 8. Experimental Design 8 Water Avoidance stress + Vehicle Water Avoidance stress + Isotype F(ab’)2 Control Water Avoidance stress + Anti-CGRP F(ab’)2 SHAM + Vehicle SHAM + Isotype F(ab’)2 Control SHAM + Anti-CGRP F(ab’)2
  • 9. Experimental Design 9 Unpredictable + Vehicle Unpredictable + Isotype F(ab’)2 Control Unpredictable + Anti-CGRP F(ab’)2 Odor only + Vehicle Odor only + Isotype F(ab’)2 Control Odor only + Anti-CGRP F(ab’)2
  • 10. Results 10 1. Pharmacokinetics of Anti-CGRP F(ab’)2
  • 11. Results 11 2. Anti-CGRP F(ab’)2 Inhibits Colonic Hypersensitivity Induced by WAS without an Effect on Colonic Compliance Two way ANOVA test followed by Bonferroni post test.
  • 12. Results 12 3. Anti-CGRP F(ab’)2 Inhibits Thoracolumbar Spinal Cord Signaling Induced by CRD in Rats Previously Exposed to WAS 0 1 2 3 4 5 Vehicle Isotype F(ab')2 Anti-CGRP F(ab')2 T10-12 SHAM WAS Average Number of pERK-IR cells/section *** *** #### ***P < 0.001 and ****P < 0.0001 compared with WAS 1 VEH; #### P < 0.0001 compared WAS 1 isotype F(ab’)2 control.
  • 13. Results 13 3. Anti-CGRP F(ab’)2 Inhibits Thoracolumbar Spinal Cord Signaling Induced by CRD in Rats Previously Exposed to WAS Average Number of pERK-IR cells/section 0 1 2 3 4 5 Vehicle Isotype F(ab')2 Anti-CGRP F(ab')2 T13-L1 SHAM WAS #### ***P < 0.001 and ****P < 0.0001 compared with WAS 1 VEH; #### P < 0.0001 compared WAS 1 isotype F(ab’)2 control. **** ****
  • 14. Results 14 4. Rodent Model of ELS: Behavioral Activation, Odor Preference, and Weaning Weights
  • 15. Results 15 4. Rodent Model of ELS: Behavioral Activation, Odor Preference, and Weaning Weights 0 1 2 3 4 5 Odor Only Unpredictable Odor Preference Odor Choices 0 10 20 30 40 50 Odor only Unpredictable Weaning Weights Weight (gm)
  • 16. Results 16 5. Anti-CGRP F(ab’)2 Inhibits Colonic Hypersensitivity in Adult Animals after Neonatal Unpredictable ELS without an Effect on Colonic Compliance
  • 17. Results 17 6. Anti-CGRP F(ab’)2 Inhibits Thoracolumbar Spinal Cord Signaling Induced by CRD in Rats Previously Exposed to Neonatal Unpredictable ELS 0 2 4 6 8 Vehicle Isotype F(ab')2 Anti-CGRP F(ab')2 T10-12 Odor only Unpredictable Average Number of pERK-IR cells/section **** **** #### ***P < 0.001 and ****P < 0.0001 compared with WAS 1 VEH; #### P < 0.0001 compared WAS 1 isotype F(ab’)2 control.
  • 18. Results 18 6. Anti-CGRP F(ab’)2 Inhibits Thoracolumbar Spinal Cord Signaling Induced by CRD in Rats Previously Exposed to Neonatal Unpredictable ELS 0 1 2 3 4 5 Vehicle Isotype F(ab')2 Anti-CGRP F(ab')2 T13-L1 Odor only Unpredictable Average Number of pERK-IR cells/section **** *** #### ***P < 0.001 and ****P < 0.0001 compared with WAS 1 VEH; #### P < 0.0001 compared WAS 1 isotype F(ab’)2 control.
  • 19. Discussion 19 To evaluate whether antibody mediated peripheral blockade of CGRP prevented stress induced visceral pain in adulthood. Repeated WAS Unpredictable ELS Peripheral CGRP Colonic hypersensitivity Neuronal activation Dorsal horn of the thoracolumbar spinal cord CGRP signalling ELS-induced visceral hypersensitivity
  • 20. Conclusion 20 IBS : Multifaceted disorder Stress early life adulthood F(ab’)2 fragment antibody Targeting CGRP inhibits stress-induced colonic hypersensitivity Stimulus-evoked spinal ERK1/2 phosphorylation Regulation of persistent visceral pain