UAB 9405 
March 16, 1994 
PAGE - 11 
5.0 DRUG INFORMATION 
The polynucleotide vaccine strategy employs the direct, intramuscular injection of naked 
plasmid DNA. The human CEA genes have been inserted into a simplified eukaryotic 
expression vector which utilizes the CMV major intermediate early promoter/enhancer to 
stimulate expression of the CEA. The plasmids are derived from the commercially 
available eucaryotic expression vector pcDNA3 (Invitrogen). The plasmid structure 
contains the cytomegalovirus early promoter/enhancer and the bovine growth hormone 
polyadenylation signal flanking a polylinker for insertion of heterologous open reading 
frames. The pcDNA3 plasmid has been modified by removal of sequences encoding the 
SV40 origin of replication and the neomycin resistance gene. Additionally, gene 
sequences encoding kanamycin resistance have been added. A detailed plasmid map is 
enclosed. The plasmid DNAs are grown in the E. Coli host strain IQ. Purification is by 
anion exchange chromatograpy utilizing Qiagen columns followed by extraction with 
Triton X-1 14 to remove endotoxin. The identity of the plasmid is verified by restriction 
endonuclease analysis. Purity of prepared DNA is validated by gel analysis, restriction 
endonuclease analysis and functional analysis for elicitation of immunity in a murine 
model of polynucleotide vaccination. Purified plasmid DNA for polynucleotide 
vaccination will be stored in lyophilized form in vials of 0.5 mg or 1.0 mg in sterile 
vacuoles. At the time of administration, the DNA will be resuspended in 1 ml sterile 
saline for injection and the appropriate volume administered at each dose level. The IND 
for this preparation will be obtained prior to trial initiation. 
6.0 TREATMENT PLAN 
6.1 Informed Consent 
Prior to entry into the study, the protocol will be discussed with the patient and 
signed informed consent obtained. 
6.2 Vaccination Strategy and Schedule 
6.2.1 Overall Approach 
Patients will be placed sequentially on study in groups of three patients 
with progressively increasing intensity of immunization. The decision to 
progress to the next dose level for patients receiving a single vaccination 
will be based on lack of acute toxicity following two weeks of follow-up 
and eight weeks of follow-up for the patients receiving three 
immunizations. 
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Recombinant DNA Research, Volume 19 
