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THE EFFECT OF SAQUINAVIR ON THE RATE OF METABOLISM OF
MIDAZOLAM
6. AUTHOR(S)
CAPT TODD BRIAN G
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7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES)
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UNIFORMED SERVICES UNIV OF HEALTH SCIENC
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THE DEPARTMENT OF THE AIR FORCE
AFIT/CIA, BLDG 125
FY99-616
2950 P STREET
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ABSTRACT
Standard Form 298 (Rev. 2-89) (EG)
Prescribed by ANSI Std. 239.18
(HI Redacted]
Designed using Perform Pro, WHS/DIOR, Oct 94
FROM : USUHS/GSN PHONE NO. : 3815995437 Oct. 0B 1999 12:50PM P3
THE EFFECT OF S AQUINAVIR ON THE RATE OF METABOUSM OF
MDDAZOLAM
Brian Todd, Capt, USAF, NC
APPROVED
cCardiy, Capt, PHS, CRNA, Ph.P, FAAN, Chair of Committee Approval Da~
Chair of Committee Approval bate
Date
AApppprroovvaall ;T
äüff9
Eugene LMine, B.B.A., M.P.A., Ph.D, Member Approval,
APPROVED:
F. O. AbdeHah, EdD., ScD., RN, FAAN, Dean
ABSTRACT
Midazolam (MDZ) is used extensively for sedation by clinicians including anesthesia
providers. There have been reports that its effects have been enhanced in the presence of
other drugs with negative patient outcomes. The P450 mixed function oxidase system
contains an enzyme subfamily known as CYP3A. These enzymes have been identified as
primarily responsible for the metabolism of MDZ. Protease inhibitors such as saquinavir
may inhibit some P450 isoforms. The metabolism of MDZ in the presence of protease
inhibitors is presently unclear. This study examines the effect of saquinavir on the rate of
metabolism of MDZ. Human liver microsomes were incubated with MDZ with and
without saquinavir. The ratio between MDZ's major metabolite oc-hydroxy MDZ and the
internal standard lorazepam were obtained using high performance liquid
chromotography (HPLC). These ratios were then compared to those results in the
presence of saquinavir. The incubation concentrations of MDZ were similar to
therapeutic concentrations of (0.5uM, luM, 3uM, 6uM, 12uM) and saquinavir (O.OuM,
0.3uM, l.OuM, 3.0uM and lO.OuM). MDZ was inhibited 56.73% ±5.63. The Ki was
determined to be 3.4 uM. The interaction exhibited properties of mixed inhibition.
These results showed that the rate of metabolism of MDZ was statistically significantly
decreases in the presence of saquinavir (p<05).
Key Words: protease inhibitor saquinavir midazolam metabolism
_„,„„. 20000112 053
Curriculum Vitae
Name: Brian George Todd.
Degree and date to be conferred: Master of Science in Nursing (6 Octoberl999).
fPII Redacted]
Secondary Education: Voorhees HS, Glen Gardner, New Jersey.
Collegiate Instituitions Attended: Dates Degree Date of Degree
USUHS, Bethesda, Maryland 1997-1999 MSN 1999.
Kutztown University, Kutztown Pa. 1989-1992 BSN 1992.
Raritian Community College, Northbranch NJ 1982-1986 ASN 1986.
Incarnate Word College, San Antonio TX 1993-1994.
Major: Nursing
Professional Publications: Poster for USUHS Research day May of 1998. The effects
of Indinavir on the rate of midazolam.
Professional Positions held:
1993-1997 Wilford Hall Medical Center, San Antonio, TX.
1990-1993 Lehigh Valley Hospital Center, Allentown, PA.
1987-1990 Easton Nursing Center, Easton, PA.
1987-1991 Kimberly Quality Care, Allentown, PA.
1986-1987 Medical Personal Pool, Allentown, PA .
li
DISCLAIMER STATEMENT
Department of Defense
This work was supported by the Uniformed Services University of the Health
Sciences. The opinions or assertions contained herein are the private opinions of the
author and are not to be construed as official or reflecting the views of the Department of
Defense or the Uniformed Services University of the Health Sciences.
111
COPYRIGHT STATEMENT
The author hereby certifies that the use of any copyrighted material in the thesis
entitled:
"THE EFFECT OF SAQUINAVIR ON THE RATE OF
METABOLISM OF MIDAZOLAM"
beyond brief excerpts is with the permission of the copyright owner, and will save
and hold harmless the Uniformed Services University of the Health Sciences from
any damage which may arise from such copyright violations.
IV
ABSTRACT
Midazolam (MDZ) is used extensively for sedation by clinicians including anesthesia
providers. There have been reports that its effects have been enhanced in the presence of
other drugs with negative patient outcomes. The P450 mixed function oxidase system
contains an enzyme subfamily known as CYP3A. These enzymes have been identified as
primarily responsible for the metabolism of MDZ. Protease inhibitors such as saquinavir
may inhibit some P450 isoforms. The metabolism of MDZ in the presence of protease
inhibitors is presently unclear. This study examines the effect of saquinavir on the rate of
metabolism of MDZ. Human liver microsomes were incubated with MDZ with and
without saquinavir. The ratio between MDZ's major metabolite ct-hydroxy MDZ and the
internal standard lorazepam were obtained using high performance liquid
chromotography (HPLC). These ratios were then compared to those results in the
presence of saquinavir. The incubation concentrations of MDZ were similar to
therapeutic concentrations of (0.5jxM, luM, 3uM, 6^iM, 12uM) and saquinavir (0.0|iM,
0.3J4M, and lO-OpM). MDZ was inhibited 56.73% ±5.63. The Ki was
I.OMM, 3.0MM
determined to be 3.4 The interaction exhibited properties of mixed inhibition.
MM.
These results showed that the rate of metabolism of MDZ was statistically significantly
decreases in the presence of saquinavir (p<.05).
Key Words: protease inhibitor saquinavir midazolam metabolism
THE EFFECT OF SAQUINAVIR ON THE RATE OF
METABOLISM OF MIDAZOLAM
Brian G. Todd, Capt, BSN
THESIS
Presented to the Graduate School of Nursing Faculty of
The Uniformed Services University of the Health
Sciences in Partial Fulfillment of the
Requirements for the
Degree of
MASTER OF SCIENCE
UNIFORMED SERVICES UNIVERSITY OF THE HEALTH SCIENCES
October 1999
vi
Ifll Redacted]
PREFACE
This study was conducted to provide information regarding the kinetic interaction
between midazolam and saquinavir. Midazolam is a frequently used medication and
could have potentially catastrophic problems if there is an unrecognized interaction with
saquinavir. This study explores this interaction in vitro.
Vll
DEDICATION
To my family, Bridget, Lauren, and Natalie. Thank you for your support during this
process and taking the time to hear me ramble on about what I am doing. I also want to
extend my most grateful thanks to Svetlana Chershnikova Ph.D. for leading me through
the process experiment and to Dr. Louis Cantilena for believing in me, that I would be
able to undertake this study.
vm
