Modulation of immunostimulatory activity of immunostimulatory ...

Stein and Cheng. Antisense oligonucleotides as therapeutic agents— is the bullet really magical? Science. Aug. 20, 1993;261(5124)1100412.*

Weiner GJ, Liu HM, Wooldridge JE, Dahle CE, Krieg AM. Immunostimulatory oligodeoxynucleotides containing the CpG motif are effective as immune adjuvants in tumor antigen imrnunization. Proc Natl Acad Sci U S A. Sep. 30, 1997;94(20)110833-7.* Reese (1978) “The Chemical Synthesis of Oligo- and PolyNucleotides By The Phosporotriester Approach,” Tetrahedron 3413143-3179.

Beaucage et al. (1981) “Deoxynucleoslde Phosphorarnidites—A New Class of Key Intermediates for Deoxypolynucleotide Synthesis” Tetrahedron Lett. 2211859-1862.

Connolly et al. (1984) “Synthesis and Characterization of an Octanucleotide Containing the EcoRI Recognition Sequence With a Phosphorothioate Group At The Cleavage Site,” Biochemistry 2313443.

Agrawal et al. (1987) “Oligodeoxynucleotise Methylphosphonatesz Synthesis and Enzymic Degradation,” Tetrahedron Lett. 28(31)3 5394542.

Jager et al. (1988) Oilgonucleotide N-Alkylphosphoroarnidatesz Synthesis and Binding to Polynucleotides, Biochemisty 2717237. Agrawal et al. (1988) Oligodeoxynucleoside Phosphoroamidates and Phosporothioates As inhibitors of Human Immunodeficiency Virus. Proc. Natl. Acad Scl. USA 8517079-7083.

Zon et al. (1991) “Phosphorothioate Oligorculeotides” Oligoncleotides and Analogues: A Practical Approach pp. 87-108. Kuramoto et al. (1992) “Oligonucleotide Sequences Required for Natural Killer Cell Activation,” Jpn. ./'. Cancer Res. 8311128-1131. Crooke (1993) “An Overview of Pro gress in Antisense Therapeutics” 8Antisense &Nucl. AcidDrugDev. 115-122 CRC Press. Boca Raton, Florida.

Zon (1993) “Protocols for Oliognucleotides and Analogs,” Methods in Molecular Biology vol. 20. pp. 165-189.

Pisetsky at al. (1994)“Stimulation of Murine Lymphocyte Proliferation by a Phosphorothioate Oligonucleotide With Antisense Activity for Herpes Simplex Virus” 54 Life Sci. 101.

Yamamoto et al. (1994) “Lipofection of Synthetic Oligodeoxyribonucleotide Having a Palindromic Sequence of AACGTT to Murine Spenocytes Enhances Interferon Production and Natural Killer Activity,” 38 Microbiol. Immunol. 831.

Agrawal et al. (1995) “Modified Oligonucleotides as Therapeutic and Diagnostic Agents” Curr Opin.Biotechnol. 6112-19.

Krieg et al. (1995) “CpG Motifs in Bacterial DNA Trigger Direct B-Cell Activation,” Nature 3711546-549.

Klinman et al.(1996) “CpG Motifs Present in Bacterial DNA Rapidly Induce Lymphocytes to Secrete Interleukin 6. Interleukin 12, and Interferon y,” 93 Proc. Natl. Acad. Sci. USA 2879.

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Primary Examiner — Zachariah Lucas

Assistant Examiner — Michelle S Horning

(74) Attorney, Agent, or Firm — Wayne A. Keown; Furman Gregory Deptula

(57) ABSTRACT

The invention relates to the therapeutic use of oligonucleotides or oligonucleotide analogs as imrnunostimulatory agents in imrnunotherapy applications. The invention provides methods for enhancing the immune response caused by imrnunostimulatory oligonucleotide compounds.

7 Claims, 53 Drawing Sheets

OTHER PUBLICATIONS

Liang et al. (1996) “Activation of Human B Cells by Phosphorothioate Oligodeoxynucleotides,” J'. Clin Invest. 98:1119-1129.

Zhao et al. (1996) “Effect of Different Chemically Modified Oligodeoxynucleotides on Immune Stimulation,” Biochem. Pharm. 5 1: 173- 182.

Chu et al. (1997) “CpG Oligodeoxynucteolides Act As Adjuvants That Switch on T Helper 1 (Thl) Immunity” 186 J'. Exp. Med. 1623. Dunford et al. (1997) “Antisense 97: Targeting the Molecular Basis of Disease” (Nature Biotechnology) Conference Abstract, pp. 40. Sparyvasser et al. (1997) “Macrophages Sense Pathogens Via DNA Motifs: Induction of Tumor Necrosis Factor-01-Mediated Shock” 27 Eur". J'. Immunol. 1671.

Zhao et al. (1997) “Pattern and Kinetics of Cytokine Production Following Administration of Phosphorothioate Oligonucleotides in Mice” 7 Antisense Nucleic Add Drug. Dev. 495.

McCluside et al. (1998) “Cutting Edge: CpG DNA Is A Potent Enhancer of Systemic and Mucosal Immune Responses Against Hepatitis B Surface Antigen with Intranasal Administration to Mice,” J'. Immunol. 161:4463.4466.

Moldoveanu et al.(1998) “CpG DNA. A Novel Immune Enhancer for Systemic and Mucosal Imn1uniZation With InfluenZa Virus” Vaccine 16: 1216-1224.

Sparyvasser et al. (1998) “Bacterial DNA and Imniunostimulatory CpG Oligonucleotides Trigger Maturation and ACtivation of Murine Dendritic Cells” 28 Eur". J'. Immunol. 2045.

Tokunaga et al. (1999) “How BCG Led to the Discovery of Imniunostimulatory DNA, ” 52 Jap. J'. Infect. Dis. 1.

Zhao et al. (1999) “Site of Chemical Modifications in CpG Containning Phosphorothioate Oligodeoxynucleotide Modulates Its Imniunostimulatory Activity,” Bioorg. & Med. Chem. Lett. 9:34533458.

Agrawal et al. (2000) “Antisense Therapeutics: Is It As Simple As Complementary Base Recognition,” 6 Mol. Med. Today 72.

Zhao et al. (2000) “Immuno stimulatory Activity of CpG Containning Phosphorothioate Oligodeoxynucleotide Is Modulated by Modification of a Single Deoxynucleoside,” Bioorg. & Med. Chem. Lett.

10:1051-1054.

Agrawal et al., “Antisense therapeutics”, Curr. Opin. Chem. Biol., 2:519-528,1998.

Chaix et al., “3'-3‘ Linked Oligonucleotides:Synthesis and Stability Studies ”, Biorg. & Med. Chem., 6:827-832, 1996.

Klinman, “therapeutic Applications of CpG-Containing OliGodeoxynucleotides”, Antisense & Nucl. Acid Drug Dev., 8:181184, 1998.

Yu et al., “Accessible 5'-End ofCpG-Containing . . . ”, Bioorganic & Medicinal Chemistry Lett., 10:2585-2588, 2000.

Kandimalla et al., “Effect of Chemical Modifications . . . ”, Bioorganic & Medicinal Chemistry, 9:807-813, 2001.

International Search Report (PCT APP. No. PCT/US01/30137).

* cited by examiner