VAC-3S immunotherapeutic HIV vaccine combined with ART is Immunogenic and Safe.

Phase II Initial Analysis of the IPROTECT1 Multicenter European Study.

Christine Katlama1, JK Rockstroh2, JM Gatell3, R Ho Tsong Fang4A, P-M Girard5, L Slama6, A Simon7, O Launay8, L Cotte9, J Reynes10, S Gharakhanian4B.1AP-HP Pitié-Salpetriere & Inserm UMR-S945, Paris 13, France; 2University of Bonn, Bonn, Germany; 3Hospital Clinic, Barcelona, Spain; 4AInnaVirVax, Genopole Evry, France; 5AP-HP St-

Antoine, Paris 12, France; 6AP-HP Tenon, Paris 20, France; 7AP-HP Pitié-Salpetriere, Paris 13, France; 8AP-HP Cochin-Pasteur & Inserm CIC, Paris 14, France; 9Croix Rousse Hospital,

Lyon, France; 10University Hospital, Montpellier, France; 4BCambridge Innovation Center, Cambridge MA, USA, & the IPROTECT1 Study Group.

We have developed an innovative immunotherapy based on a highly specific and conserved motif,

called 3S, located in the gp41 HIV-1 protein. This highly pathogenic motif induces expression of

NKp44L, the cellular ligand of an activating NK receptor (NKp44), rendering uninfected CD4+ T cells

sensitive to NK lysis1,5,7,13 (Figure 1). NKp44L expression is strongly correlated with both the decline

of CD4 cell count2,6,8,9,11,15,18, and additional effects linked to the pathogenecity (ie. T-cell activation,

inflammation, and apoptosis of CD4+ T cells). These data were observed in vitro after 3S

stimulation1,5,7, ex-vivo in HIV-infected patients1, and in vivo in SHIV-infected macaques4. A

protection against CD4 depletion and chronic immune activation was demonstrated in SHIV-

infected macaques immunized with a 3S vaccine3,10. Non clinical data3,10,14,17, as well as the safety

and immunogenicity results19-25 of the phase I/IIa clinical trial IVVAC-3S/P1 have been presented.

Final analysis of this first-in-man study showed immunological effects with increased CD4/CD8 ratio,

due to significant decrease in CD8 % and increase in CD4 % and virological effects with a significant

decrease in HIV reservoir that was correlated to anti-3S antibody titers26.

We present here preliminary data on safety and immunogenicity of the phase IIa IPROTECT1 two

months after the end of enrollment.

Prospective, randomized, placebo-controlled, double-blind, 3-step study in Europe,

assessing immunotherapeutic properties of VAC-3S at 16, 32, 64 mg with 3 IM base

immunizations at 4 week intervals and 3 maintenance boosters in the 16, 32 µg arms.

Ninety HIV ART suppressed patients with 200-349 and 350-500 CD4 cells/mm3

planned, 40 % and 60 % of the patients respectively. Primary endpoint is

immunogenicity measured by total anti-3S Ig at week 12 measured by ELISA.

Secondary endpoints include: T lymphocyte activation/differentiation, HIV DNA,

inflammatory biomarkers.

VAC-3S is confirmed safe and immunogenic in HIV-infected

patients with CD4 counts between 200 and 500 CD4+ T

cells/mm3. The current schedule of administration permits

sustained levels of anti-3S antibodies. Further analysis will be

conducted in order to confirm biological effects on T cell

homeostasis and HIV reservoir observed during the phase I/IIa.

Abstract MOPEA038

Figure 1 : Depletion of uninfected CD4+ T cell by 3S.Steps ① & ②: The gp41 3S motif of HIV-1 binds to its specific receptor (gC1qR) on CD4+ T cells.Step ③: This interaction induces NKp44L expression at the surface of non-infected CD4+ T cells.Steps ④ to ⑥: NKp44L/NKp44 interaction provokes NK-mediated cytotoxicity.

INTRODUCTION

Table 1 : Demography. 86 patients were enrolled in the study in 13 investigational centers inFrance, Spain and Germany: 36 patients in the low CD4 strata and 50 patients in the high CD4strata; 18 women and 68 men.

SAFETY

Figure 4 : Immunogenicity. The first step of inclusion permitted to evaluate the safety of the32 µg dose, the second step of inclusion of the 64 µg dose. During these two steps, the minimumrequired interval between two inclusions was of 48h. The third step of inclusion enrolled theremnant of the patient population without timing limitation between inclusions. Total Anti-3S Igwere measured by ELISA. Arbitrary Units are reported on a log10 scale for each patient. Blue verticalbars: VAC-3S/placebo administration. Temporary threshold of 30 A.U. above which immunologicaland virological effects were demonstrated during the phase I/IIa is represented by the grey area.Results are blinded.

IMMUNOGENICITY

Table 2 : SafetyAll related (possibly related, probably related and related) Treatment Emergent Adverse Events arereported by severity for 86 patients who received 291 VAC-3S/Placebo Administrationscorresponding to a total of 582 injections (an administration = 1 injection in each arm formethodology constraints).

Figure 3 : Sample size / Randomisation. The total sample size of the study to evaluate thenull hypothesis for the primary endpoint is estimated at 90 patients assuming an overall type Ierror of 2.5 %, one-sided test, with the power of at least 90 %. The assumptions regarding humoralresponses are 0% in the placebo group, 52 % in the 16 µg group, 62 % in the 32 µg group and 72 %in the 64 µg group.DEMOGRAPHY

METHODS

CONCLUSION

REFERENCES

ACKNOWLEDGEMENTS

Tota

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(A

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Un

its)

Weeks

First Step of Inclusion:6 patients : 32 µg3 patients : 16 µg2 patients : placebo

Second step of inclusion:6 patients : 64 µg3 patients : 16 or 32 µg2 patients : placebo

Third step of inclusion:55 patients : 16 µg, 32 µg,

64 µg or placebo

Figure 2 : Study Plan

1. Vieillard V et.al. NK cytotoxicity against CD4+ T cells during HIV-1 infection: A gp41 peptide induces the expression of an NKp44 ligand.Proc. Natl. Acad. Sci USA, 102:10981-10986. 2. Vieillard V et.al. 2006. Specific adaptive humoral response against a gp41 motif inhibits CD4 T-cells sensitivity to NK lysis during HIV-1 infection. AIDS 20:1795-18043. Vieillard V et.al 2008. A new vaccine strategy against AIDS: A HIV gpg41 peptide immunization prevents NKp44L expression and CD4+ T cell depletion in SHIV-infected monkeys. Proc.Natl. Acad. Sci. USA, 105:2100-4.4. Vieillard V et.al 2008. CCR5 or CXCR4 use influences the relationshipbetween CD4 cell depletion, NKp44L expression and NK cytotoxicity in SHIV-infected macaques. AIDS, 22:185–192.5. Fauster-Bovendo H et.al 2009. HIV escape from natural killer cytotoxicity : nef inhibits NKp44L expression on CD4+ T cells. AIDS 23:1077–1087.6. Vieillard V et.al 2010. Specific Phenotypic and Functional Features of Natural Killer Cells From HIV-Infected Long-Term Nonprogressors and HIV Controllers. J. Acquir. Immune Defic. Syndr. 53:564-5737. Fausther-Bovendo H., et.al 2010. HIV gp41 Engages gC1qR on CD4+ T Cells to Induce the Expression of an NK Ligand through the PIP3/H2O2 Pathway. PLoS Pathogens, 6:1-13. 8. M. Curriu, et.al 2010. Viremic HIV Infected Individuals with High CD4 T Cells and Functional Envelope Proteins Show Anti-gp41 Antibodieswith Unique Specificity and Function. 2012. PLos ONE, 7:1-12.9. Vieillard V et.al 2012. Specific anti-gp41 antibodies predict HIV-1 disease progression. 2012. J. Acquir. Immune Defic. Syndr. 61:403-405.10.Vieillard, V et.al An HIVgp41 vaccine protects CD4 central memory cells. 2012. Vaccine, 30:6883-891.11. Sennepin A. et.al NKp44L expression on CD4+ T cells is associated

with impaired immunologic recovery in HIV-infected patients underhighly active antiretroviral therapy. 2013. AIDS, 27:1857-1866.12.C. Petitdemange, et.al A Single Amino-acid Change in a HighlyConserved Motif of gp41 Elicits Virus Neutralization and Protectsagainst CD4 Depletion. CID, First Published Online May 21, 2013.13.F. Baychelier, et.al Identification of a cellular ligand for the naturalcytotoxicity recepetor NKp44. 2013. Blood, 122: 2935-2942.14.Vincent Vieillard et.al .October 2009, AIDS Vaccine 2009, Paris, France. Poster 19-25. A vaccine model to prevent the depletion of uninfected bystander CD4 cells during HIV infection.15.Vincent Vieillard et.al. March 2011, 15th international workshop on HIV observational databases, Prague, Czech Republic. Specificadaptative humoral response against a gp41 motif is associated withdelayed HIV disease progression. 16. J. Crouzet et.al. January 2013, Workshop on HIV cure and eradication : a feasible option ?, Milan, ItalyPotential role of anti-3S antibodies in an HIV medical cure strategy ? 17.R. Ho Tsong Fang et.al. January 2013, Société Française d’Immunologie, 8ème colloque du club de Vaccinologie, Institut Pasteur, Paris, France. Dose-Ranging for Development of VAC-3S, a novel HIV Immunotherapeutic Vaccine.18.A. Sennepin et.al. March 2013, 20th Conference on Retroviruses and Opportunistic Infections, Atlanta, USA. Poster 312. NKp44L Expression on CD4+ T cells Is Associated with Impaired Immunological Recovery in HIV-1+ Patients Under HAART 19. C. Katlama et.al. March 2013, 20th Conference on Retroviruses and Opportunistic Infections, Atlanta, USA; Poster 392; A randomized, placebo-controlled, double-blind, phase I/IIa dose escalation study of an HIV immunotherapeutic vaccine, VAC-3S, directed to the gp41 3S motif of HIV-1.20.Christine Katlama et.al., May 2013, 30 Years of HIV Science, Imagine

the Future, Pasteur Institute, Paris, France. Poster 145. ExperimentalEvidence and Potential Immunotherapeutic Applications of Vaccine-Induced Antibodies Against 3S, a Highly Conserved Motif of gp41, in HIV-1-infected Patients Treated with Antiretroviral Therapy.21.R. Calin, et.al., June 2013, 14es Journées Nationales d’Infectiologie, Clermont-Ferrand, France. Etude de phase I/IIa de VAC-3S : Un vaccin immunothérapeutique /immunoprotecteur, en complément des antirétroviraux, au cours de l’infection VIH. 22.S Gharakhanian et.al., October 2013, AIDS Vaccine 2013, Barcelona, Spain. VAC-3S, an Immunoprotective HIV Vaccine Directed to the 3S Motif of gp41, in Patients Receiving ART: Safety, Dose & Immunization Schedule Assessment. 23.C. Katlama et.al. June 2014, AFRAVIH, Montpellier, France. Etude de phase I/IIa de VAC-3S :un vaccin immunothérapeutique/immunoprotecteur, en complément des antirétroviraux, au cours de l’infection VIH. [French]24.H Bodilis et.al., June 2014, 15èmes journées nationales d’infectiologie, Bordeaux, FranceVAC-3S, un nouveau vaccin immunothérapeutique de l’infection VIH-1 : résultats de la phase I. [French]25.R. Ho Tsong Fang et.al., September 2014, 4th International Conference on vaccines and vaccination, Valencia, Spain. VAC-3S vaccine, a novel approach to the therapeutic management of HIV Infection. Overview of phase I and phase II clinical vaccine development programs. 26.Ho Tsong Fang et. al. July 2015, Towards an HIV Cure Symposium, Vancouver, Canada. VAC-3S, a safe Immunotherapeutic HIV Vaccine decreased total HIV DNA and increased CD4/CD8 ratio: Phase I Final Results. Abstract 3630.

The authors would like to thank the patients and their entourage; Principal investigators: Pr. Gilles Pialoux, Pr. Alain Devidas, Pr. Laurence Weiss, Pr. Yasdan Yasdanpanah, Pr. Gerd Fätkenheuer; Co-investigators and staff of clinical centers: at Hospitals Cochin, Dr Aziza Chermak, Pitié-Salpêtrière (Infectious and Tropical Diseases ; Internal Medicine Units), Dr Nadia Valin, Saint-Antoine, Tenon, Pompidou, Paris; Corinne Brochier, Hospices Civils de Lyon, Lyon; CHU de Montpellier, Montpellier, Hôpital Sud-Francilien, Corbeil, FRANCE; Internal Medicine, Medical Faculty, Köln University, Dr. Christoph Boesecke, Pavlos Kokordelis, Department of General internal Medicine, Medical Faculty, University of Bonn, GERMANY, Pr Felipe Garcia, Hospital Clinic, University of Barcelona, Barcelona, SPAIN; all InnaVirVax team, Evry, France (D. Batéjat, N. Baran, A. Beurdeley, C. Blaison, M. Galaup, G. Heinrich, S. Lebel-Binay, D. Lucas, M. Nizou-Marcu, M. Sicard, T. Varela); B. Orlandini, PhinC Development, Evry, France; Delphine Aubert, Antoinette ANGER, Effi-Stat, Paris, FRANCE; Maud Reynierand staff of Eurofins Optimed, Grenoble, FRANCE.

Patient Disposition Strata CD4 200-349 Strata CD4 350-500 All Treated

Median [Range] N=36 N=50 N=86

Gender H/F 28 / 8 40 / 10 68 / 18

Age (years) 48.5 [23-59] 48.0 [27-61] 48.0 [23-61]

Weight (kg) 69.0 [49-119] 76.2 [57-107] 73.0 [49-119]

Body Mass Index (kg/m2) 22.5 [16.3-33.4] 25.0 [18.7-33.5] 24.0 [16.3-33.5]

Patients who received three administrations 28/36 36/50 64/86

Nadir CD4 (cells/mm3) 131.5 [10-410] 176.0 [21-375] 159.5 [110-410]

Baseline CD4 counts (cells/mm3) 287.8 [189-428] 416.3 [259-596] 364.3 [189-596]

Baseline CD8 counts (cells/mm3) 587.0 [230.5-1172.5] 610.3 [137.5-1960] 602.3 [137.5-1960.0]

Baseline CD4 % 23.9 [12.5-42.0] 28.4 [16.0-43.0] 27.0 [12.5-43.0]

Baseline CD8 % 47.8 [26.5-66.0] 40.8 [13.0-78.0] 43.3 [13.0-78.0]

Baseline Ratio CD4/CD8 0.51 [0.21-1.23] 0.68 [0.22-2.90] 0.63 [0.21-2.90]

Source : PROTECT1 2015-05-29/Analysis/Programs/IAS 2015.sas

Baseline = Mean (Screening - D0)

All Related Treatment-emergent

Adverse Events

N=86 Event Subject (%) Event Subject Event Subject Event Subject Event Subject

Local 127 40 (47 %) 44 15 (18 %) 7 3 (4 %) 0 0 (0 %) 178 43 (50 %)

Pain 52 26 (31 %) 25 10 (12 %) 3 2 (3 %) 80 29 (34 %)

Sensitivity 45 17 (20 %) 10 5 (6 %) 2 1 (2 %) 57 22 (26 %)

Induration 12 4 (5 %) 2 2 (3 %) 14 6 (7 %)

Swelling 2 2 (3 %) 2 1 (2 %) 4 3 (4 %)

Pruritus 2 1 (2 %) 2 1 (2 %)

Oedema 6 2 (3 %) 1 1 (2 %) 7 3 (4 %)

Erythema 10 6 (7 %) 4 4 (5 %) 14 9 (11 %)

General 57 19 (23 %) 29 12 (14 %) 2 2 (3 %) 0 0 (0 %) 88 24 (28 %)

Pain 25 10 (12 %) 9 3 (4 %) 34 11 (13 %)

Asthenia 9 7 (9 %) 5 3 (4 %) 1 1 (2 %) 15 11 (13 %)

Headache 4 2 (3 %) 6 4 (5 %) 1 1 (2 %) 11 6 (7 %)

Fever 5 2 (3 %) 3 2 (3 %) 8 3 (4 %)

Nausea 4 3 (4 %) 1 1 (2 %) 5 4 (5 %)

Myalgia 6 4 (5 %) 4 3 (4 %) 10 6 (7 %)

Vertigo 4 4 (5 %) 1 1 (2 %) 5 4 (5 %)

Others 10 7 (9 %) 8 6 (7 %) 1 2 (3 %) 0 0 (0 %) 19 12 (14 %)

NUMBNESS OF UPPER ARM 2 1 (2 %) 1 1 (2 %) 3 2 (3 %)

ARTHRALGIA MULTIPLE 1 1 (2 %) 1 1 (2 %)

CPK INCREASE 1 1 (2 %) 1 1 (2 %)

HERPES LABIALIS 2 1 (2 %) 2 1 (2 %)

PRURITUS FACIAL 1 1 (2 %) 1 1 (2 %)

TENDINITIS 1 1 (2 %) 1 1 (2 %)

HYPERESTHESIA 1 1 (2 %) 1 1 (2 %)

ACNE 1 1 (2 %) 1 1 (2 %)

ADENOMIALGY 1 1 (2 %) 1 1 (2 %)

AXILLARY ADENOPATHY 1 1 (2 %) 1 1 (2 %)

FLU SYNDROME 1 1 (2 %) 1 1 (2 %)

ALOPECIA 1 1 (2 %) 1 1 (2 %)

DIARRHEA 1 1 (2 %) 1 1 (2 %)

TOXICODERMA 1 1 (2 %) 1 1 (2 %)

STINGING 2 1 (2 %) 2 1 (2 %)

TOTAL 285 51 (59 %)

Grade 1

Mild

Grade 2

Moderate

Grade 3

Severe

Grade 4

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