Clinical experience with recombinant human FSH

1
Clinical experience with recombinant human FSH
Zeev Shoham, M.D.
Department of Obstetrics and Gynecology, Kaplan
Hospital, Rehovot, Israel
rec-hFSH
2
Clinical experience with recombinant human FSH
Zeev Shoham, M.D.
Department of Obstetrics and Gynecology, Kaplan
Medical Center , Rehovot, Israel
rec-hFSH
3
Human FSH

• Glycoprotein hormone (a and b subunits).
• a subunit composed of 92 amino acids and carries
  2 carbohydrate moieties.
• b subunit composed of 111 amino acids and carries
  2 carbohydrate moieties.
• FSH molecule exists in many different (iso)forms
  due to differences in carbohydrate moieties.
• Carbohydrate moieties are important in
  determining half-life of FSH and therefore
  modulates in-vivo biological action.

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The Production of a Simple Protein by Recombinant
DNA Techniques
Insertion into a specific vector
Messenger RNA
Complementary-DNA
Reverse transcriptase
Transfection of the cell
Cell producing a protein
Manufacture of the protein
Kaplan Medical Center, Rehovot, Israel
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DNA The Building Block of Life
DNA carries the instructions to make all the
proteins necessary for the normal function of the
cell. A gene is a region of DNA that encodes one
function (protein).
DNA library
Genes books
Kaplan Medical Center, Rehovot, Israel
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Recombinant DNA production became possible due to
four major discoveries
Kaplan Medical Center, Rehovot, Israel
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Tools
Restriction enzymes Scissors
Ligases enzymes Glue
Reverse transcriptase Synthesis DNA directly
from mRNA Complementary DNA
Kaplan Medical Center, Rehovot, Israel
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Tools
Bacterial plasmids
Viral vectors
Kaplan Medical Center, Rehovot, Israel
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Expression of hFSH in Mammalian Cells
Chinese Hamster Ovary (CHO) cell line used
Very well characterized cell line, used for the
production of other bio-engineering proteins.
Perform post-translation modification with high
efficiency.
Capable of proper assembly of subunits and
glycosilation.
Kaplan Medical Center, Rehovot, Israel
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Expression of hFSH in CHO Cells
a
Incorporation into host cell Chromosome
Nucleus
Transfection
Transcription
b
a
mRNA produced
Golgi apparatus
Maturation
Translation
ER
Secretion
b
a
Mature hFSH molecule
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Production of r-hFSH
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The Three-Dimensional Model Structure of hFSH
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Mean Pharmacokinetic Parameters after a single
Injection of 300 or 150 IU, IM, of rec-hFSH
Peak recFSH concentration C max (IU/L/h)
Time of occurrence T max (h)
Elimination half life T 1/2
(h)
300
150
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First established pregnancy using rec-hFSH and
GnRH-a for IVF-ET treatment
hCG 10,000
amps.
OC
4
ET - 4
rec-hFSH
1
18
E
Mid-luteal phase
2
14
Buserelin 0.2 mg
8
Menses
21
7
2
Germond et al. Lancet 1992
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rec-FSH vs. U-FSH - a prospective multicenter
study
No. of oocytes
Centers
Kaplan Medical Center, Rehovot, Israel
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rec-FSH vs. U-FSH - a prospective multicenter
study
Out et al. Hum Reprod. 1995
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rec-FSH vs. U-FSH - a prospective multicenter
study
Out et al. Hum Reprod. 1995
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rec-FSH vs. U-FSH - a prospective multicenter
study
Recombinant human FSH study group Fertil Steril
1995
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rec-FSH vs. U-FSH - a prospective multicenter
study
Recombinant human FSH study group Fertil Steril
1995
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conclusion

• It is clear today that the use of recombinant
  human gonadotropins is expected to provide
• better batch-to-batch consistency,
• steady supply,
• a purified compound,
• with high specific activity, which accounts for
  gt99 of the preparations protein content.

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conclusion
Recombinant FSH is well tolerated. No drug
related adverse reaction was noted. No serum
antirecombinant FSH antibody was detected.
Mannaerts et al. Fertil Steril 1993 Shoham et al.
Fertil Steril 1993 Schoot et al. J Clin
Endocrinol Metab. 1992
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conclusion
Its efficacy in ovulation induction and ovarian
stimulation for assisted reproduction has been
clearly demonstrated.
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conclusion
There is no doubt that recombinant gonadotropins
produced by genetic engineering technology are
here to stay and will represent an important
treatment modality in various fertility
disturbances.
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Rec-FSH vs. hMG in PCOS patients
(Severe OHSS in previous cycle)
Foll.gt16 mm
1/2 amp
14 D
hCG 5000 IU
Aboulghar et al. Fertil Steril 1996
Kaplan Medical Center, Rehovot, Israel
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Rec-FSH for ovarian stimulation for IVF-ET
gt17 mm
hCG 10,000
19 cycles
52 cycles
Menses
Strowitzki et al. Hum Reprod. 1995
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FSH Isoforms
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FUTURE

• Separation of isoforms
• long half-life
• short half-life
• PCO moment to moment control
• Fusion molecules FSH with hCG
• Development of Orally Active Gonadotropin
  Mimetics

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Rec-FSH for ovarian stimulation for IVF-ET
Menses
Hendon et al. Hum Reprod. 1995