Post on 11-Mar-2020
Transnasal Delivery of a Gonadotropin-releasing Hormone Analog In Miniature
Swine
Mark Santos (Microbiology Major)Department of Biochemistry and Molecular Biology
Robert Gyurik, Zhengmao Li, Carl Reppucci, Scott Kavanaugh, Roger Wells and Stacia A. Sower
Vertebrate 1 2 3 4 5 6 7 8 9 10Mammal pGlu His Trp Ser Tyr Gly Leu Arg Pro GlyNH2
Guinea Pig pGlu Tyr Trp Ser Tyr Gly Val Arg Pro GlyNH2Chicken - I pGlu His Trp Ser Tyr Gly Leu Gln Pro GlyNH2
Rana pGlu His Trp Ser Tyr Gly Leu Trp Pro GlyNH2Seabream pGlu His Trp Ser Tyr Gly Leu Ser Pro GlyNH2
Salmon pGlu His Trp Ser Tyr Gly Trp Leu Pro GlyNH2Medaka pGlu His Trp Ser Phe Gly Leu Ser Pro GlyNH2Catfish pGlu His Trp Ser His Gly Leu Asn Pro GlyNH2Herring pGlu His Trp Ser His Gly Leu Ser Pro GlyNH2
Chicken -II pGlu His Trp Ser His Gly Trp Tyr Pro GlyNH2Dogfish pGlu His Trp Ser His Gly Trp Leu Pro GlyNH2
Lamprey - III pGlu His Trp Ser His Asp Trp Lys Pro GlyNH2Lamprey - I pGlu His Tyr Ser Leu Glu Trp Lys Pro GlyNH2Whitefish pGlu His Trp Ser Tyr Gly Met Asn Pro GlyNH2
Vertebrate Family of Peptides
Mihael Freamat
What is Gonadotropin-Releasing Hormone (GnRH)?
Strategy for Developing GnRH Analogs
pGlu1 - His2 - Trp3 - Ser4 - Tyr5 - Gly6 - Leu7 - Arg8 - Pro7 - Gly10NH2
Agonist(Stimulator)
Antagonist(Inhibitor)
D-Amino Acid(Especially Bulky Aromatics)
D-Amino Acid(Especially Basics)
D-Ala
EthylamideD-Amino Acid
(Especially Aromatics) NαMe-Leu
Identification of novel GnRHs and Receptors can contribute to the development of novel GnRH therapeutics.
How Is GnRH Used Medically10,000 Analogs Developed
http://perso.wanadoo.fr/pharma.rocheville/contraception_nouveautes.htm
Westside Pregnancy Resource Center
FertilizationContraceptionCancer Therapy
http://www.news-medical.net/images/breast%20cancer%20cell.jpg
Breast cancer cell
Why Miniature Swine?Swine have been shown to have similar mucosal membrane properties
to humans making them an ideal test subject (Bollen et al., 2000).
ObjectiveTo determine if the GnRH analog DAla6Pro9NEt can successfully be
delivered via a transnasal application.
GnRH Complex
Biological Effect
Measure GnRH & LH
MethodsCatheter Implantation to 4 Miniature Swine
Plasma Collection for GnRH Analysis
Serum Collection for LH Analysis
GnRH Extracted From Plasma
Samples Sent to Dr. Richard Barb
Samples Assayed Using a Specific
Radioimmunoassay
Samples Assayed Using a Specific
Radioimmunoassay
Data Analyzed
Dr. Roger Wells and Van Gould
This study was completed using 4 miniature swine aged for 4 months. They were cared for according to IACUC requirements.
Animals
The TreatmentsPig 1: 0% CPE 0% GnRH Analog (Control)
Pig 2: 2% CPE 1% GnRH Analog
Pig 3: 0% CPE 1% GnRH Analog
Pig 4: 0% CPE 2% GnRH Analog
Sampling Times
Week 1 Days 1+2
Week 2 Days 1+2
Week 1 Days 3+4
Week 2 Days 3+4
0 Hours30 Min60 Min90 Min
180 Min24 Hours
0 Hours30 Min60 Min90 Min
180 Min360 Min
24 Hours
Week 1 Days 1+2
Week 2 Days 1+2
Week 1 Days 3+4
Week 2 Days 3+4
0 Hours30 Min60 Min90 Min
180 Min24 Hours
0 Hours30 Min60 Min90 Min
180 Min360 Min
24 Hours
Sampling Times
GnRH - Week 1 Days 1 + 2
These results show a significant increase in GnRH concentration in the blood using the Bentley Formulation with GnRH
BB B B
BB
J
J
J
J
J
J
H
H
H
H
H
H
FF
F
FF
F
0 150 300 450 600 750 900 1050 1200 13500
200
400
600
800
1000
1200
1400
GnR
H p
g/0.
1ml
Sampling Time (Minutes)
B ControlJ 2%CPE 1%GnRHH 0%CPE 1%GnRHF 0%CPE 2%GnRH
Week 1 Days 1+2
GnRH - Week 1 Days 3+4
These results show a significant increase in GnRH concentration in the blood using the Bentley Formulation with GnRH
BB
BB
B B
BJ
J
J
J
J
J JH
H
H
H HH
HF
F
F
F
FF F
0 150 300 450 600 750 900 1050 1200 13500
50
100
150
200
250
GnR
H p
g/0.
1ml
Sampling Time (Minutes)
B ControlJ 2%CPE 1%GnRHH 0%CPE 1%GnRHF 0%CPE 2%GnRH
Week 1 Days 3+4
GnRH - Week 2 Days 1+2
These results show a significant increase in GnRH concentration in the blood using the Bentley Formulation with GnRH
B B B B B BJ
J
J
J
JJH
H
H
H
H H
F F FF
F F0 150 300 450 600 750 900 1050 1200 1350
0
20
40
60
80
100
120
GnR
H (P
g/0.
1ml)
Time (Minutes)
B 0% CPE 0% GnRHJ 2% CPE 1% GnRHH 0% CPE 1% GnRHF 0% CPE 2% GnRH
Week 2 Days 1 + 2
GnRH - Week 2 Days 3+4
These results show a significant increase in GnRH concentration in the blood using the Bentley Formulation with GnRH
B B B B B B BJ
JJ
J
J
JJ
H
H HH
H H H
FF
FF
F F F0 150 300 450 600 750 900 1050 1200 1350
0
50
100
150
200
250
300
350
GnR
H (P
g/0.
1ml)
Time (Minutes)
B 0% CPE 0% GnRHJ 2% CPE 1% GnRHH 0% CPE 1% GnRHF 0% CPE 2% GnRH
Week 2 Days 3 + 4
Summary
These studies showed that GnRH combined with the Bentley formulation
was successfully administered into the pig following transnasal application.
Implications
The results from this project can be eventually used by pharmaceutical industries
in development of drug applications.