National Toxicology Program

National Toxicology Program
https://ntp.niehs.nih.gov/go/16217

IMM98009: Immunotoxicity of Saquinavir in Female B6C3F1 Mice

The following abstract presents results of a study conducted by a contract laboratory for the National Toxicology Program (NTP). The findings have not been peer reviewed and were not evaluated in accordance with the levels of evidence criteria established by the NTP on March 2009 (see http://ntp.niehs.nih.gov/ntp/htdocs/levels/09-3566%20NTP-ITOX-R1.pdf). The findings and conclusions for this study should not be construed to represent the views of the NTP or the US Government.

FINAL REPORT
Saquinavir (CAS No. 149845-06-7)

NTP Study Number: IMM98009

Saquinavir (SQV) is a protease inhibitor, targeted to interfere late in the viral life cycle during cleavage of the final viral protein element. The protease inhibitors bind to the catalytic site of the viral protease enzyme that plays a critical role in cleaving the large polyproteins of the immature virus. The protease inhibitors represent an alternate treatment for AIDS patients and, since the protease inhibitors act at a totally different site in the viral cycle, a combination treatment with nucleoside analogues may be effective. Reasons for concern are seen in the potential cross resistance between different protease inhibitors and the potential for drug interactions since the protease inhibitors are metabolized in the liver and some are potent inhibitors of cytochrome p4501.

Saquinavir was nominated to the NTP for toxicological evaluation and was selected for immunotoxicity studies by the project officer. Thus, the purpose of these studies was to determine the potential effects of SQV on the immune system.

The study reported herein was conducted in female B6C3F1 mice. The animals were treated twice daily for 28 days by oral gavage. The in-life phase of the study was conducted between 13 February 1998 and 03 September 1998. Saquinavir was prepared weekly as a solution in the vehicle 0.5% methylcellulose.

The baseline toxicology studies are summarized in Table ES-1. In this study of orally administered SQV at a total dose of 300, 600, and 1200 mg/kg/day for 28 days, there were no effects on body weight or on the rate of weight gain. There were few notable changes among the standard toxicological parameters that were assessed. The reticulocyte count was significantly decreased by 12% at the high dose (1200 mg/kg) level.

Table ES-2 summarizes the immunology studies. A statistically significant increase of 23% was observed at the high dose level of SQV in the absolute number of T cells. The immature CD4+CD8+ cells, when evaluated as absolute values, showed an increase of 46% and 92% at the middle and high dose levels, respectively. The humoral immune response was significantly increased by 41% for AFC/106 spleen cells at the high dose level; however, there was no effect on the anti-sRBC serum titer. There was no effect on cell-mediated immunity when evaluated by the mixed leukocyte (MLR) response. However, cytotoxic T-cell activity showed decreases ranging from 25% to 35% at the low dose level for effector-to-target ratios of 6.25:1, 12.5:1 and 25:1. Overall, Natural Killer cell activity was not altered in SQV-treated mice.

Table ES-3 summarizes the two host resistance studies that were conducted. Overall, exposure to SQV did not affect host resistance to Listeria monocytogenes; however, a statistically significant decrease in host resistance to Listeria monocytogenes was observed at the 600 mg/kg dose level for Challenge B. Exposure to SQV did not alter host resistance to the B16F10 melanoma tumor, a model system which evaluates effects on innate immunity, including natural killer cells and macrophages, as well as the cytotoxic T-cell population of cell-mediated immunity.

Table ES-1
SUMMARY TABLE FOR TOXICOLOGY STUDIES
SQV-28-1M-PO
Parameter Overall Effect Specific Results Maximum Effect Dose Comment
Body Weight No Effect



Day 1
No Effect


Day 8
No Effect


Day 15
No Effect


Day 22
No Effect


Day 29
No Effect


Body Weight Changes No Effect



Day 8-1
No Effect


Day 15-1
No Effect


Day 22-1
No Effect


Day 29-1
No Effect


Pathology




Gross




Pathology




Histopathology




Organ Weights No Effect



Liver
No Effect


Spleen
No Effect


Lungs
No Effect


Thymus
No Effect


Kidney
No Effect


Hematology No Effect



RBCs
No Effect


Hemoglobin
No Effect


Hematocrit
No Effect


MCV
No Effect


MCH
No Effect


MCHC
Increase 1% 600 mg/kg
Reticulocytes
Decrease 12% 1200 mg/kg
Platelets
No Effect


Leukocytes
No Effect




Table ES-2
SUMMARY TABLE FOR IMMUNOLOGY STUDIES
SQV-28-1M-PO
Parameter Overall Effect Specific Results Maximum Effect Dose Comment
Surface Makers (Absolute Values)

No Effect



Ig+
No Effect


CD3+
Increase 23% 1200 mg/kg
CD4+CD8-
No Effect


CD4-CD8+
No Effect


CD4+CD8+
Increase 92% 1200 mg/kg
NK1.1+CD3-
No Effect


Mac-3+
No Effect


IgM Humoral Immune Response to Sheep Erythrocytes

Increased



IgM AFC/106Cells
Increase 41% 1200 mg/kg
Serum Titer
No Effect


Mixed Leukocyte Response

No Effect



Responders Only
No Effect


Responders + Stimulators
No Effect


Total Cells x 107
No Effect


Cytotoxic T Lymphocyte Response

No Effect



6:1
Decrease 35% 300 mg/kg
12.5:1
Decrease 28% 300 mg/kg
25:1
Decrease 25% 300 mg/kg
NK Cell Activity





No Effect



200:1
No Effect


100:1
Decrease 20% 300 mg/kg
50:1
Decrease 27% 300 mg/kg
25:1
Decrease 27% 300 mg/kg
12.5:1
Decrease 31% 300 mg/kg
6.25:1
Decrease 40% 1200 mg/kg


Table ES-3
SUMMARY TABLE FOR HOST RESISTANCE STUDIES
SQV-28-1M-PO
Parameter Overall Effect Specific Results Maximum Effect Dose Comment
Listeria monocytogenes No Effect Decrease 8% mortality 600 mg/kg Challenge B
B16F10 No Effect No Effect


Report Date: October 2004

NTP is located at the National Institute of Environmental Health Sciences, part of the National Institutes of Health.