Protocol 031

OUTLINE OF PROTOCOL 031

A PHASE I SAFETY AND IMMUNOGENICITY TRIAL OF THE FACILITATED HIV-1 gag-pol DNA VACCINE (APL-400-047, APOLLON, INC.) GIVEN INTRAMUSCULARLY BY NEEDLE AND SYRINGE OR BIOJECTOR 2000^® NEEDLE-FREE JET INJECTION SYSTEM IN HIV-1 UNINFECTED ADULT VOLUNTEERS

Subjects: 52 HIV-1 uninfected, healthy adult volunteers aged 18-60 years old with lower risk for HIV-1 infection (AVEG categories A or B).

Schema: Amended 7/98 to include Group V; Amendment pending to boost consenting volunteers in Groups II-IV with ALVAC-HIV (vCP205) [Pasteur Merieux Connaught]

ORIGINAL STUDY

Group	N^*	Delivery System	Immunization Schedule in Months
Group	N^*	Delivery System	0	1	2	6
I	8/2	Needle/Syringe	100 µg	100 µg	100 µg	100 µg
II	8/2	Needle/Syringe	300 µg	300 µg	300 µg	300 µg
III	8/2	Biojector	300 µg	300 µg	300 µg	300 µg
IV	8/2	Needle/Syringe	1000 µg	1000 µg	1000 µg	1000 µg
TOTAL	32/8	.	* Within each group of 10 volunteers, 8 will receive APL-400-047, formulated with bupivacaine as a facilitating agent, 2 will receive control preparation (bupivacaine carrier alone)

AMENDMENT II

Group

N

Delivery System

Immunization Schedule in Months

0

1

2

6

V

10/2 ^**

Needle/Syringe

3000 µg

3000 µg

3000 µg

3000 µg

TOTAL 42/10
.

** Within the 12 volunteers in Group V, 10 volunteers, will receive APL-400-047, formulated with bupivacaine as a facilitating agent, 2 will receive control preparation (bupivacaine carrier alone). An injection of 1.5 ml will be given into the deltoid muscle of each arm.

AMENDMENT III and IV: ADDITIONAL BOOSTER IMMUNIZATIONS, GROUPS II - V

Original Study Group

N

DNA Original Vaccine Dose

DNA + Boost Immunization Schedule (Months) ^#

0

1

2

II

8/2

300 µg N/S ^##

DNA (IM)

vCP205

vCP205

III

8/2

300 µg Biojector

IV

8/2

1000 µg N/S

V 10/2 3000 µg N/S DNA (IM) vCP 1452 vCP 1452

^# Consenting volunteers will receive an additional dose of DNA (or control) followed by two monthly canarypox or placebo boosts

^## N/S: needle/syringe

DNA boost: 1000 µg given IM (Groups II, III, IV); 3000 µg given IM (Group V)
vCP205: ALVAC-HIV vCP205, approximate dose of 10^6.7TCID₅₀ ,given IM
vCP1452: ALVAC-HIV vCP1452, approximate dose of 10^6.7TCID₅₀ ,given IM
Placebo: Placebo ALVAC (diluent for vCP205 and vCP1452)

ACCRUAL AND IMMUNIZATIONS COMPLETED FOR GROUPS I-V PER ORIGINAL STUDY, AMENDMENTS II AND III (AMENDMENT IV FOR ADDITIONAL BOOSTER IMMUNIZATIONS FOR GROUP V IS PENDING)

Product Description: The APL 400-047 DNA plasmid vaccine expresses the human immunodeficiency virus type-1 core structural proteins, and partially deleted nonfunctional reverse transcriptase and integrase proteins, derived from the HIV-1 strain HXB2 and inserted into the GENEVAX^® DNA plasmid backbone. APL 400-047 is formulated with the facilitating agent bupivacaine, an amide-type local anesthetic agent. [Apollon (presently Wyeth Lederle Vaccines)]

ALVAC-HIV vCP205 is a recombinant canarypox vector into which the following genes have been inserted: HIV-1 envelope gp120 (strain MN) linked to the transmembrane portion of HIV-1 gp41 (strain LAI) and the HIV-1 LAI genes encoding the entire gag protein and a portion of the pol sequence, sufficient to evoke the protease portion. [Pasteur-Merieux/Connaught]

Time Period: 18 months, extended to 5 months past the first Amendment III immunization; Groups I-IV: first volunteer enrolled 07/16/97 and the last on 10/30/97; Group V: first volunteer enrolled 11/30/98 and the last on 02/23/99; Amendment III boosts began 08/99.

Clinical Sites: University of Alabama at Birmingham, University of Rochester, University of Washington, Vanderbilt University

Study Chair: Mark Mulligan, University of Alabama at Birmingham

INCLUSION CRITERIA

Standard inclusion criteria, with the addition of:

CPK < 2 x institutional upper normal limit
ANA < 1:80
Negative for anti-dsDNA antibodies

Amendments III and IV:

Allergy to egg products or neomycin (used to prepare ALVAC vaccines).

EXCLUSION CRITERIA

Standard exclusion criteria, with the addition of:

Hypersensitivity to bupivacaine or other amide-type anesthetics (i.e., lidocaine, mepivacaine, etc.)

STUDY GOALS

To perform a randomized, double-blind, controlled Phase I trial of the facilitated HIV-1 gag-pol DNA vaccine, APL-400-047 (Apollon, Inc.) in 40 lower risk (AVEG risk groups A or B), healthy, HIV-1 uninfected adult volunteers aged 18-60 years.
To evaluate the safety and tolerability in humans of the APL-400-047 when administered intramuscularly by needle and syringe at a dose of 100 µg.
If initially safe and well tolerated at 100 µg: to evaluate the safety and tolerability of the facilitated APL-400-047 when administered intramuscularly either by needle and syringe or Biojector at a dose of 300 µg.
If 300 µg dose delivered by needle and syringe is safe and well tolerated: to evaluate the safety and tolerability of the facilitated APL-400-047 when administered at a dose of 1,000 µg given by needle and syringe.
To evaluate the immunogenicity of the APL-400-047 formulated with bupivacaine when administered at doses of 100, 300, or 1,000 µg by needle and syringe injection or 300 µg via Biojector delivery. The HIV-1 gag-pol specific immune responses to be assessed include: binding antibodies, lymphocyte proliferation, cytotoxic T-lymphocytes, and ancillary assays.
Amendment II: To evaluate the tolerability, safety and immunogenicity of a dose of 3,000 µg in an additional group of volunteers.
Amendment III: To evaluate the tolerability, safety and immunogenicity of an additional dose of 1000 µg of the APL-400-047 DNA vaccine, followed by two doses of live recombinant canarypox ALVAC-HIV vCP205 given one and two months later, in volunteers who received four previous doses of 300 or 1000 µg of APL-400-047.
Amendment IV: To evaluate the tolerability, safety and immunogenicity of an additional dose of 3000 µg of the APL-400-047 DNA vaccine, followed by two doses of live recombinant canarypox HIV vCP1452 given one and two months later, in volunteers who received four previous doses of 3000 µg of APL-400-047.

Original Study Group	N	DNA Original Vaccine Dose	DNA + Boost Immunization Schedule (Months) ^#
			0	1	2
II	8/2	300 µg N/S ^##	DNA (IM)	vCP205	vCP205
III	8/2	300 µg Biojector
IV	8/2	1000 µg N/S
V	10/2	3000 µg N/S	DNA (IM)	vCP 1452	vCP 1452
^# Consenting volunteers will receive an additional dose of DNA (or control) followed by two monthly canarypox or placebo boosts ^## N/S: needle/syringe			DNA boost: 1000 µg given IM (Groups II, III, IV); 3000 µg given IM (Group V) vCP205: ALVAC-HIV vCP205, approximate dose of 10^6.7TCID₅₀ ,given IM vCP1452: ALVAC-HIV vCP1452, approximate dose of 10^6.7TCID₅₀ ,given IM Placebo: Placebo ALVAC (diluent for vCP205 and vCP1452)