(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2004/0102389 A1
`(43) Pub. Date:
`May 27, 2004
`Pavco et al.
`
`US 20040102389A1
`
`(54) NUCLEIC ACID-MEDIATED TREATMENT
`OF DISEASES OR CONDITIONS RELATED
`TO LEVELS OF VASCULAR ENDOTHELIAL
`GROWTH FACTOR RECEPTOR (VEGF-R)
`
`(75) Inventors: Pamela Pavco, Lafayette, CO (US);
`James McSWiggen, Boulder, CO (US);
`Dan Stinchcomb, Ft. Collins, CO (US);
`Jaime Escobedo, Alamo, CA (US);
`Julian Kim, Shaker Heights, OH (US);
`Daniel Lindner, Shaker Heights, OH
`(Us)
`Correspondence Address:
`MCDONNELL BOEHNEN HULBERT &
`BERGHOFF
`300 SOUTH WACKER DRIVE
`SUITE 3200
`CHICAGO, IL 60606 (US)
`
`(73) Assignee: Ribozyme Pharmaceuticals, Inc.
`
`(21) Appl. No.:
`
`10/287,949
`
`(22) Filed:
`
`Nov. 4, 2002
`
`Related US. Application Data
`
`(63) Continuation-in-part of application No. 10/138,674,
`?led on May 3, 2002, Which is a continuation-in-part
`of application No. 09/870,161, ?led on May 29, 2001,
`Which is a continuation-in-part of application No.
`
`09/708,690, ?led on Nov. 7, 2000, Which is a con
`tinuation-in-part of application No. 09/371,722, ?led
`on Aug. 10, 1999, noW Pat. No. 6,534,872, Which is
`a continuation-in-part of application No. 08/584,040,
`?led on Jan. 11, 1996, noW Pat. No. 6,346,398.
`
`(60) Provisional application No. 60/005,974, ?led on Oct.
`26, 1995.
`
`(30)
`
`Foreign Application Priority Data
`
`Oct. 25, 1996 (WO) ......................... .. PCT/US96/17480
`May 29, 2002 (WO) ......................... .. PCT/US02/17674
`
`Publication Classi?cation
`
`.. A61K 48/00; C12N 5/08
`(51) Int. Cl.7
`(52) US. Cl. ........................... .. 514/44; 435/372; 435/375
`
`(57)
`
`ABSTRACT
`
`The present invention relates to nucleic acid molecules such
`as riboZymes, DNAZymes, short interfering RNA (siRNA),
`short interfering nuleic acid (siNA), and antisense Which
`modulate the synthesis, expression and/or stability of an
`mRNA encoding one or more receptors of vascular endot
`helial groWth factor, such as ?t-1 (VEGFRl) and/or KDR
`(VEGFR2). Nucleic acid molecules and methods for the
`inhibition of angiogenesis and treatment of cancer and other
`conditions associated With VEGF-R are provided, optionally
`in conjunction With other therapeutic agents such as inter
`ferons.
`
`1
`
`MTX1049
`
`

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`Patent Application Publication May 27, 2004 Sheet 1 0f 40
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`Patent Application Publication May 27, 2004 Sheet 2 0f 40
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`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 3 0f 40
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`Patent Application Publication May 27, 2004 Sheet 4 0f 40
`
`US 2004/0102389 A1
`
`Figure 4: Hepatitis Delta Virus Ribozyme
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`

`Patent Application Publication May 27, 2004 Sheet 5 0f 40
`
`US 2004/0102389 A1
`
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`Patent Application Publication May 27, 2004 Sheet 6 0f 40
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`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 7 0f 40
`
`US 2004/0102389 A1
`
`Figure 7: Anti-FLT Ribozyme-Mediated
`Inhibition of VEGF Binding to FLT Receptor
`(Human Microvascular Endothelial Cells)
`
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`Patent Application Publication May 27, 2004 Sheet 8 0f 40
`
`US 2004/0102389 A1
`
`4
`Figure 8: Anti-KDR Ribozyme-Mediated
`Inhibition of VEGF Binding to KDR Receptor
`(Human Microvas'cular Endothelial Cells)
`
`'Ribozymes ‘
`
`Ribozyrnes
`
`9
`
`

`

`Patent Application Publication May 27, 2004 Sheet 9 0f 40
`
`US 2004/0102389 A1
`
`Figure 9: Speci?city of Anti-FLT Ribozyme
`Mediated Inhibition of VEGF Binding
`(Human Microvascular Endothelial Cells) ‘
`
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`

`Patent Application Publication May 27, 2004 Sheet 10 0f 40
`
`US 2004/0102389 A1
`
`Figure 10: Anti-KDR Antisense-Mediated Inhibition of
`Human aortic endothelial cell (HAEC) Proliferation
`
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`Patent Application Publication May 27, 2004 Sheet 12 0f 40
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`Patent Application Publication May 27, 2004 Sheet 13 0f 40
`
`US 2004/0102389 A1
`
`Figure 11C: Cleavage of ?t-l RNA by 1358 HH
`Ribozymes
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`
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`Patent Application Publication May 27, 2004 Sheet 14 0f 40
`
`US 2004/0102389 A1
`
`Figure 11D : Cleavage of ?t-l RNA by 4229 HH
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`Patent Application Publication May 27, 2004 Sheet 16 of 40
`
`US 2004/0102389 A1
`
`Figure 12BzAnti-flt-1 Ribozyme-Mediated Inhibition
`of Cell Proliferation
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`Patent Application Publication May 27, 2004 Sheet 17 0f 40
`
`US 2004/0102389 A1
`
`Figure l3zAnti-KDR Ribozyme-Mediated Inhibition
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`Patent Application Publication May 27, 2004 Sheet 18 0f 40
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`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 19 0f 40
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`Patent Application Publication May 27, 2004 Sheet 20 0f 40
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`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 21 of 40
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`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 22 0f 40
`
`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 23 0f 40
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`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 24 0f 40
`
`US 2004/0102389 A1
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`
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`Patent Application Publication May 27, 2004 Sheet 25 0f 40
`
`US 2004/0102389 A1
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`
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`Patent Application Publication May 27, 2004 Sheet 26 0f 40
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`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 28 of 40
`
`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 29 of 40
`
`US 2004/0102389 A1
`
`
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`Patent Application Publication May 27, 2004 Sheet 30 0f 40
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`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 35 of 40
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`US 2004/0102389 A1
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`Patent Application Publication May 27, 2004 Sheet 40 0f 40
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`US 2004/0102389 A1
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`US 2004/0102389 A1
`
`May 27, 2004
`
`NUCLEIC ACID-MEDIATED TREATMENT OF
`DISEASES OR CONDITIONS RELATED TO
`LEVELS OF VASCULAR ENDOTHELIAL
`GROWTH FACTOR RECEPTOR (VEGF-R)
`
`[0001] This patent application is a continuation-in-part of
`Pavco et al., US. Ser. No. 10/138,674, filed May 3, 2002,
`entitled “Enzymatic Nucleic Acid-Mediated Treatment of
`Ocular Diseases or Conditions Related to Levels of Vascular
`
`Endothelial Growth Factor Receptor (VEGF-R)” which is a
`continuation-in-part of Pavco et al., US. Ser. No. 09/870,
`161, filed May 29, 2001, which is a continuation-in-part of
`Pavco et al., US. Ser. No. 09/708,690, filed Nov. 7, 2000,
`which is a continuation-in-part of Pavco et al., US. Ser. No.
`09/371,722, filed Aug. 10, 1999, which is a continuation-
`in-part of Pavco et al., US. Ser. No. 08/584,040, filed Jan.
`11, 1996, which claims the benefit of Pavco et al., US. S No.
`60/005,974, filed on Oct. 26, 1995, all which are entitled
`“Method and Reagent for Treatment of Diseases or Condi-
`tions Related To Levels of Vascular Endothelial Growth
`
`Factor Receptor”. Each of these applications is hereby
`incorporated by reference herein in its entirety including the
`drawings and tables.
`
`BACKGROUND OF THE INVENTION
`
`[0002] This invention relates to methods and reagents for
`the treatment of diseases or conditions relating to the levels
`of expression of vascular endothelial growth factor (VEGF)
`receptor(s).
`
`[0003] The following is a discussion of relevant art, none
`of which is admitted to be prior art to the present invention.
`
`[0004] VEGF, also referred to as vascular permeability
`factor (VPF) and vasculotropin,
`is a potent and highly
`specific mitogen of vascular endothelial cells (for a review
`see Ferrara, 1993 Trends Cardiovas. Med. 3, 244; Neufeld et
`al., 1994 Prog. Growth Factor Res. 5, 89). VEGF induced
`neovascularization is implicated in various pathological
`conditions such as tumor angiogenesis, proliferative diabetic
`retinopathy, hypoxia-induced angiogenesis,
`rheumatoid
`arthritis, psoriasis, wound healing and others.
`
`[0005] VEGF, an endothelial cell-specific mitogen, is a
`34-45 kDa glycoprotein with a wide range of activities that
`include promotion of angiogenesis, enhancement of vascu-
`lar-permeability and others. VEGF belongs to the platelet-
`derived growth factor (PDGF) family of growth factors with
`approximately 18% homology with the A and B chain of
`PDGF at the amino acid level. Additionally, VEGF contains
`the eight conserved cysteine residues common to all growth
`factors belonging to the PDGF family (Neufeld et al., supra).
`VEGF protein is believed to exist predominantly as disul-
`fide-linked homodimers; monomers of VEGF have been
`shown to be inactive (Plouet et al, 1989 EMBO J. 8, 3801).
`
`[0006] VEGF exerts its influence on vascular endothelial
`cells by binding to specific high-affinity cell surface recep-
`tors. Covalent cross-linking experiments with 125I-labeled
`VEGF protein have led to the identification of three high
`molecular weight complexes of 225, 195 and 175 kDa
`presumed to be VEGF and VEGF receptor complexes (Vais-
`man et al., 1990 J. Biol. Chem. 265, 19461). Based on these
`studies VEGF-specific receptors of 180, 150 and 130 kDa
`molecular mass were predicted. In endothelial cells, recep-
`tors of 150 and the 130 kDa have been identified. The VEGF
`
`receptors belong to the superfamily of receptor tyrosine
`kinases (RTKs) characterized by a conserved cytoplasmic
`catalytic kinase domain and a hydrophylic kinase sequence.
`The extracellular domains of the VEGF receptors consist of
`seven immunoglobulin-like domains that are thought to be
`involved in VEGF binding functions.
`
`[0007] The two most abundant and high-affinity receptors
`of VEGF are fit-1 (fms-like tyrosine kinase) cloned by
`Shibuya et al., 1990 Oncogene 5, 519 and KDR (kinase-
`insert-domain-containing receptor) cloned by Terman et al.,
`1991 Oncogene 6, 1677. The murine homolog of KDR,
`cloned by Mathews et al., 1991, Proc. Natl. Acad. Sci, USA,
`88, 9026, shares 85% amino acid homology with KDR and
`is termed as fik-1 (fetal liver kinase-1). Recently it has been
`shown that the high-affinity binding of VEGF to its receptors
`is modulated by cell surface-associated heparin and heparin-
`like molecules (Gitay-Goren et al., 1992 J. Biol. Chem. 267,
`6093).
`
`[0008] VEGF expression has been associated with several
`pathological states such as tumor angiogenesis, several
`forms of blindness, rheumatoid arthritis, psoriasis and oth-
`ers. Following is a brief summary of evidence supporting the
`involvement of VEGF in various diseases:
`
`1) Tumor angiogenesis: Increased levels of VEGF
`[0009]
`gene expression have been reported in vascularized and
`edema-associated brain tumors (Berkman et al., 1993 J.
`Clini. Invest. 91, 153). Amore direct demostration of the role
`of VEGF in tumor angiogenesis was demonstrated by Jim
`Kim et al., 1993 Nature 362,841 wherein, monoclonal
`antibodies against VEGF were successfully used to inhibit
`the growth of rhabdomyosarcoma, glioblastoma multiforme
`cells in nude mice. Similarly, expression of a dominant
`negative mutated form of the flt-1 VEGF receptor inhibits
`vascularization induced by human glioblastoma cells in
`nude mice (Millauer et al., 1994, Nature 367, 576).
`
`2) Ocular diseases: Aiello et al., 1994 New Engl. J.
`[0010]
`Med. 331, 1480, showed that the ocular fluid of a majority
`of subjects suffering from diabetic retinopathy and other
`retinal disorders, contains a high concentration of VEGF.
`Miller et al., 1994Am. J. Pathol. 145, 574, reported elevated
`levels of VEGF mRNA in subjects suffering from retinal
`ischemia. These observations support a direct role for VEGF
`in ocular diseases.
`
`3) Psoriasis: Detmar et al., 1994 J. Exp. Med. 180,
`[0011]
`1141 reported that VEGF and its receptors were over-
`expressed in psoriatic skin and psoriatic dermal microves-
`sels, suggesting that VEGF plays a significant role in pso-
`r1as1s.
`
`Immunohistochemistry
`4) Rheumatoid arthritis:
`[0012]
`and in situ hybridization studies on tissues from the joints of
`subjects
`suffering from rheumatoid arthritis
`show an
`increased level of VEGF and its receptors (Fava et al., 1994
`J. Exp. Med. 180, 341). Additionally, Koch et al., 1994 J.
`Immunol. 152, 4149, found that VEGF-specific antibodies
`were able to significantly reduce the mitogenic activity of
`synovial tissues from subjects suffering from rheumatoid
`arthritis. These 25 observations support a direct role for
`VEGF in rheumatoid arthritis.
`
`5) Autosomal dominant polycystic kidney disease
`[0013]
`(ADPKD): ADPKD is the most common life threatening
`hereditary disease in the USA. It affects about 1:400 to
`
`42
`
`42
`
`

`

`US 2004/0102389 A1
`
`May 27, 2004
`
`1:1000 people. Approximately 50% of people with ADPKD
`develop renal failure. ADPKD accounts for about 5-10% of
`end-stage renal failure in the United States requiring dialysis
`and renal transplantation. Several animal models, including
`the Han:SPRD rat model, mice with a targeted mutation in
`the Pkd2 gene, and congenital polycystic kidney (cpk) mice,
`closely resemble human ADPKD and present an opportunity
`to evaluate the therapeutic effect of agents that have the
`potential to interfere with one or more of the pathogenic
`elements of ADPKD. One of the features of ADPKD is
`
`angiogenesis. Angiogenesis may be necessary for growth of
`cyst cells as well as increased vascular permeability pro-
`moting fluid secretion into cysts. Proliferation of cystic
`epithelium is also a feature of ADPKD. Cyst cells in culture
`produce soluble vascular endothelial growth factor (VEGF),
`which is proven to be specific and critical for blood vessel
`formation. VEGF is also the best validated target for anti-
`angiogenesis therapies based on overwhelming genetic,
`mechanistic and animal efficacy data. However, VEGF can
`also directly stimulate proliferation of epithelial cells. VEGF
`triggers a response by interacting with cell-surface recep-
`tors. VEGFR1 has been detected in epithelial cells of cystic
`tubules but not in endothelial cells in the vasculature of
`
`cystic kidneys or normal kidneys. VEGFR2 expression is
`increased in endothelial cells of cyst vessels and in endot-
`helial cells during renal ischemia-reperfusion. It is proposed
`that inhibition of VEGF receptors with anti-VEGFR1 and
`anti-VEGFR2 (KDR) agents (eg. nucleic acid molecules of
`the invention) would attenuate cyst formation, renal failure
`and mortality in ADPKD. Anti-VEGFR2 agents (eg. nucleic
`acid molecules of the invention) would inhibit angiogenesis
`involved in cyst formation. As VEGFR1 is present in cystic
`epithelium and not in vascular endothelium of cysts, it is
`proposed that anti-VEGFR1 agents would attenuate cystic
`epithelial cell proliferation and apoptosis which would in
`turn lead to less cyst formation. Further, it is proposed that
`VEGF produced by cystic epithelial cells is one of the
`stimuli for angiogenesis as well as epithelial cell prolifera-
`tion and apoptosis. Validation assays for nucleic acid mol-
`ecules of the invention can be performed in Han:SPRD rats,
`mice with a targeted mutation in the Pkd2 gene, and cpk
`mice. The effect of anti-VEGF nucleic acids on cyst forma-
`tion and renal failure can determine the potential harmful
`role of angiogenesis in ADPKD.
`[0014]
`In addition to the above data on pathological con-
`ditions involving excessive angiogenesis, a number of stud-
`ies have demonstrated that VEGF is both necessary and
`sufficient for neovascularization. Takashita et al., 1995 J.
`Clin. Invest. 93, 662, demonstrated that a single injection of
`VEGF augmented collateral vessel development in a rabbit
`model of ischemia. VEGF also can induce neovasculariza-
`
`tion when injected into the cornea. Expression of the VEGF
`gene in CHO cells is sufficient to confer tumorigenic poten-
`tial to the cells. Kim et al., supra and Millauer et al., supra
`used monoclonal antibodies against VEGF or a dominant
`negative form of flk-1 receptor to inhibit tumor-induced
`neovascularization.
`
`[0015] During development, VEGF and its receptors are
`associated with regions of new vascular growth (Millauer et
`al., 1993 Cell 72, 835; Shalaby et al., 1993 J. Clin. Invest.
`91, 2235). Furthermore, transgenic mice lacking either of
`the VEGF receptors are defective in blood vessel formation,
`in fact these mice do not survive; fik-1 appears to be required
`for differentiation of endothelial cells, while fit-1 appears to
`
`be required at later stages of vessel formation (Shalaby et al.,
`1995 Nature 376, 62; Fung et al., 1995 Nature 376, 66).
`Thus, these receptors must be present to properly signal
`endothelial cells or their precursors to respond to vascular-
`ization-promoting stimuli.
`
`[0016] All of the conditions listed above, involve exten-
`sive vascularization. This hyper-stimulation of endothelial
`cells may be alleviated by VEGF antagonists. Thus most of
`the therapeutic efforts for the above conditions have con-
`centrated on finding inhibitors of the VEGF protein.
`
`[0017] Kim et al., 1993 Nature 362, 841 have been suc-
`cessful
`in inhibiting VEGF-induced tumor growth and
`angiogenesis in nude mice by treating the mice with VEGF-
`specific monoclonal antibody.
`
`[0018] Koch et al., 1994 J. Immunol. 152, 4149 showed
`that the mitogenic activity of microvascular endothelial cells
`found in rheumatoid arthritis (RA) synovial tissue explants
`and the chemotactic property of endothelial cells from RA
`synovial fluid can be neutralized significantly by treatment
`with VEGF-specific antibodies.
`
`[0019] Ullrich et al., International PCT Publication No.
`WO 94/11499 and Millauer et al., 1994 Nature 367, 576
`used a soluble form of fik-1 receptor (dominant-negative
`mutant) to prevent VEGF-mediated tumor angiogenesis in
`immunodeficient mice.
`
`[0020] Kendall and Thomas, International PCT Publica-
`tion No. WO 94/21679 describe the use of naturally occur-
`ing or recombinantly-engineered soluble forms of VEGF
`receptors to inhibit VEGF activity.
`
`[0021] Robinson, International PCT Publication No. WO
`95/04142 describes the use of antisense oligonucleotides
`targeted against VEGF RNA to inhibit VEGF expression.
`
`Jellinek et al., 1994 Biochemistry 33, 10450
`[0022]
`describe the use of VEGF-specific high-affinity RNA aptam-
`ers to inhibit the binding of VEGF to its receptors.
`
`[0023] Rockwell and Goldstein, International PCT Publi-
`cation No. WO 95/21868, describe the use of anti-VEGF
`receptor monoclonal antibodies to neutralize the effect of
`VEGF on endothelial cells.
`
`SUMMARY OF THE INVENTION
`
`[0024] The invention features novel nucleic acid-based
`compounds
`[e.g.,
`enzymatic nucleic
`acid molecules
`(ribozymes such as Inozyme, G-cleaver, amberzyme, zin-
`zyme), DNAzymes, antisense nucleic acids, 2-5A antisense
`chimeras, triplex forming nucleic acid, decoy nucleic acids,
`aptamers, allozymes, antisense nucleic acids containing
`RNA cleaving chemical groups (Cook et al., US. Pat. No.
`5,359,051), small interfering RNA (siRNA), small interfer-
`ing nucleic acid (siNA, Beigelman et al., US. S No. 60/409,
`293)] and methods for their use to down regulate or inhibit
`the expression of receptors of VEGF (VEGF-R such as
`VEGFR1 and/or VEGFR2).
`
`In one embodiment, the invention features the use
`[0025]
`of one or more of the nucleic acid-based compounds to
`inhibit the expression of VEGFR1 (fit-1) and/or VEGFR2
`(fik-1/KDR) receptors.
`
`43
`
`43
`
`

`

`US 2004/0102389 A1
`
`May 27, 2004
`
`In another embodiment, the present invention fea-
`[0026]
`tures a compound having Formula I: (SEQ ID NO: 20818).
`
`gsasgsusugchAuGagg
`5'
`[0027]
`cGaaAgucugB 3'
`
`ccgaaa
`
`ggc-
`
`[0028] wherein each a is 2'-O-methyl adenosine
`nucleotide, each g is a 2'-O-methyl guanosine nucle-
`otide, each c is a 2'-O-methyl cytidine nucleotide,
`each u is a 2'-O-methyl uridine nucleotide, each Ais
`adenosine, each G is guanosine, each s individually
`represents a phosphorothioate internucleotide link-
`age, U is 2'-deoxy-2'-C-allyl uridine, and B is an
`inverted deoxyabasic moiety.
`
`In another embodiment, the present invention fea-
`[0029]
`tures a compound having Formula II: (SEQ ID NO: 13488).
`
`5'-usascsasau ucU GAu Gag gcg aaa gcc Gaa
`[0030]
`Aag aca aB-3'
`
`[0031] wherein each a is 2'-O-methyl adenosine nucle-
`otide, each g is a 2'-O-methyl guanosine nucleotide, each c
`is a 2'-O-methyl cytidine nucleotide, each u is a 2'-O-methyl
`uridine nucleotide, eachAis adenosine, each G is guanosine,
`each s individually represents a phosphorothioate inter-
`nucleotide linkage, Q is 2'-deoxy-2'-C-allyl uridine, and B is
`an inverted deoxyabasic moiety.
`
`In one embodiment, the invention features a com-
`[0032]
`position comprising a compound of Formula I and/or II in a
`pharmaceutically acceptable carrier or diluent.
`
`In another embodiment, the invention features a
`[0033]
`method of administering to a cell, for example a mammalian
`cell or human cell, the compound of Formula I and/or II,
`comprising contacting the cell with the compound under
`conditions suitable for administration, for example in the
`presence of a delivery reagent. Examples of suitable delivery
`reagents include a lipid, cationic lipid, phospholipid, or
`liposome as described herein and known in the art.
`
`a
`the invention features
`In one embodiment,
`[0034]
`method of administering to a cell the compound of Formula
`I or II in conjunction with a chemotherapeutic agent com-
`prising contacting the cell with the compound and the
`chemotherapeutic
`agent under conditions
`suitable
`for
`administration.
`
`[0035] Examples of chemotherapeutic agents that can be
`combined with the compound of Formula I and/or II include
`but are not limited to S-fluoro uridine, Leucovorin, Irinote-
`can (CAMPTOSAR® or CPT—11 or Camptothecin-11 or
`Campto), Paclitaxel, or Carboplatin or
`a combination
`thereof.
`
`In another embodiment, the present invention also
`[0036]
`features a cell comprising the compound of Formula I and/or
`II, wherein the cell is a mammalian cell. For example, in one
`embodiment the mammalian cell is a human cell.
`
`a
`the invention features
`In one embodiment,
`[0037]
`method of inhibiting angiogenesis,
`for example tumor
`angiogenesis, in a subject comprising the step of contacting
`the subject with the compound of Formula I and/or II under
`conditions suitable for said inhibition. In one embodiment,
`the subject is a mammal, for example, a human.
`
`In another embodiment, the invention features a
`[0038]
`method of treatment of a subject having a condition asso-
`
`for
`ciated with an increased level of VEGF receptor,
`example, cancers such as breast cancer, lung cancer, col-
`orectal cancer, renal cancer, pancreatic cancer, or melanoma;
`Autosomal dominant polycystic kidney disease (ADPKD);
`or ocular indications such as diabetic retinopathy, or age
`related macular degeneration, comprising contacting one or
`more cells of the subject with the compound of Formula I
`and/or II, under conditions suitable for the treatment. In one
`embodiment, the subject is a human.
`
`In another embodiment, the invention features a
`[0039]
`method of treatment of a subject having an ocular condition
`associated with an increased level of a VEGF receptor, for
`example, diabetic retinopathy, or age related macular degen-
`eration, comprising contacting one or more cells of the
`subject with a nucleic acid molecule, such as an enzymatic
`nucleic acid molecule, targeted against a VEGF receptor
`RNA, e.g., a molecule according to Formula I and/or II,
`under conditions suitable for the treatment. In one embodi-
`
`ment, the subject is a human.
`
`In yet another embodiment, a method of treatment
`[0040]
`of the invention further comprises the use of one or more
`drug therapies under conditions suitable for the treatment.
`
`invention also
`the present
`In one embodiment,
`[0041]
`features a method of cleaving RNA comprising a sequence
`of VEGFR1 (fit-1) comprising contacting the compound of
`Formula I with the RNA under conditions suitable for the
`
`cleavage of the RNA, for example, where the cleavage is
`carried out in the presence of a divalent cation such as
`Mg2+.
`
`In another embodiment, the invention features a
`[0042]
`method of administering to a mammal,
`for example a
`human, the compound of Formula I and/or II comprising
`contacting the mammal with the compound under conditions
`suitable for the administration, for example, in the presence
`of a deliver