WO2007053447A2 - Compositions and methods for the treatment and prevention of fibrotic, inflammatory and neovascularization conditions - Google Patents
Compositions and methods for the treatment and prevention of fibrotic, inflammatory and neovascularization conditions Download PDFInfo
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Definitions
- the present invention relates to methods of treatments for ocular disorders using immune-derived moieties which are reactive against bioactive lipid molecules that play role in human and/or animal disease as signaling molecules.
- One particular class of signaling bioactive lipids considered in accordance with the invention is lysolipids.
- Particularly preferred signaling lysolipids are sphingosine-1 -phosphate (SlP) and the various lysophosphatidic acids (LPAs).
- SlP sphingosine-1 -phosphate
- LPAs various lysophosphatidic acids
- Antibodies against signaling lipids, and derivatives and variants thereof, can be used in the treatment and/or prevention of ocular diseases or disorders through the delivery of pharmaceutical compositions that contain such antibodies, alone or in combination with other therapeutic agents and/or treatments.
- the present invention relates to methods of decreasing or attenuating aberrant neovascularization, angiogenesis, aberrant fibrogenesis, fibrosis and scarring, and inflammation and immune responses. These processes, separately or together are involved in many diseases and conditions. These diseases or conditions may be systemic or may be relatively localized, for example to the skin or to the eye.
- Pathologic or aberrant angiogenesis/neovascularization, aberrant remodeling, fibrosis and scarring and inflammation occur in association with retinal and ocular ischemic diseases such as age-related macular degeneration (AMD), diabetic retinopathy (DR) and in retinopathy of prematurity (ROP) and other developmental disorders [Eichler et al. (2006), Curr Pharm Des, vol 12: 2645-60] as well as being a result of infections and mechanical injury to the eye [Ciulla et al. (2001), Curr Opin Ophthalmol, vol 12: 442-9 and Dart et al (2003), Eye, vol 17: 886-92].
- AMD age-related macular degeneration
- DR diabetic retinopathy
- ROP retinopathy of prematurity
- CNV Choroidal neovascularization
- AMD is used here solely for illustrative purposes in describing ocular conditions relating to aberrant angiogenesis/neovascularization, aberrant remodeling, fibrosis and scarring, and inflammation, which conditions are found in other ocular diseases and disorders as disclosed and claimed herein.
- AMD involves age-related pathologic changes [Tezel, Bora and Kaplan (2004), Trends MoI Med, vol 10: 417-20 and Zarbin (2004), Arch Ophthalmol, 122: 598-614]. Multiple theories exist but, the exact etiology and pathogenesis of AMD are still not well understood. Aging is associated with cumulative oxidative injury, thickening of Bruch's membrane and drusen formation.
- Oxidative stress results in injury to retinal pigment epithelial (RPE) cells and, in some cases, the choriocapillaris [Zarbin (2004), Arch Ophthalmol, vol 122: 598-614 and Gorin et al. (1999), MoI Vis,, vol 5: 29].
- RPE retinal pigment epithelial
- Injury to RPE likely elicits a chronic inflammatory response within Bruchs membrane and the choroid [Johnson et at (2000), Exp Eye Res,, vol 70: 441-9]. This injury and inflammation fosters and potentates retinal damage by stimulating CNV and atrophy [Zarbin (2004), Arch Ophthalmol, vol 122: 598-614 and Witmer et at (2003), Prog Retin Eye Res, vol 22: 1-29].
- BV defective and leaky blood vessels
- Wound healing arises from the choroid and invades the subretinal space through Bruchs membrane and the RPE. Wound healing responses are characterized by a typical early inflammation response, a prominent angiogenic response and tissue formation followed by end-stage maturation of all involved elements. Wound remodeling may irreversibly compromise photoreceptors and RPEs thereby, justifying the need to treat CNV with more than anti-angiogenic therapies [La Cour, Kiilgaard and Nissen (2002), Drugs Aging, vol 19: 101-33.12].
- Anti- VEGF-A therapies represent a recent, significant advance in the treatment of exudative AMD.
- the phase III VISION Trial with PEGAPT ANIB a high affinity aptamer which selectively inhibits the 165 isoform of VEGF-A, demonstrated that the average patient continues to lose vision and only a small percent gained vision [Gragoudas et at (2004), N Engl J Med, vol 351: 2805-16].
- Inhibition of all isoforms of VEGF-A (pan- VEGF inhibition) with the antibody fragment RANIBIZUMAB yielded much more impressive results [Brown et al. N Eng Med,2006 355:1432-44, Rosenfeld et al. N Eng J Med 2006355:1419-31].
- the 2 year MARINA trial and the 1 year ANCHOR trial demonstrated that approximately 40% of patients achieve some visual gain. Although these results represent a major advance in our ability to treat exudative AMD, they also demonstrate that 60% of patients do not have visual improvement. Furthermore, these patients had to meet strictly defined inclusion and exclusion criteria. The results in a larger patient population may be less robust.
- exudative AMD is comprised of vascular and extravascular components.
- the vascular component involves vascular endothelial cells (EC), EC precursors and pericytes.
- the extravascular component which volumetrically appears to be the largest component, is composed of inflammatory, glial and retinal pigment epithelium (RPE) cells and fibroblasts.
- Tissue damage can result from either component. These other aspects of the pathologic process are not addressed by current anti- VEGF treatments. Targeting additional elements of the angiogenic cascade associated with AMD could provide a more effective and synergistic approach to therapy [Spaide RF (2006), Am J Ophthalmol, vol 141: 149-156].
- Angiogenesis is an essential component of normal wound healing as it delivers oxygen and nutrients to inflammatory cells and assists in debris removal [Lingen (2001), Arch Pathol Lab Med, vol 125: 67-71].
- Progressive angiogenesis is composed of two distinct processes: Stage I: Migration of vascular ECs, in response to nearby stimuli, to the tips of the capillaries where they proliferate and form luminal structures; and Stage II: Pruning of the vessel network and optimization of the vasculature [Guo et al. (2003), Am J Pathol, vol 162: 1083-93].
- Neovascularization most often aids wound healing.
- new vessels when uncontrolled are commonly defective and promote leakage, hemorrhaging and inflammation.
- Diminishing dysfunctional and leaky BVs, by targeting pro-angiogenic GFs, has demonstrated some ability to slow the progression of AMD [Pauleikhoff (2005), Retina, vol 25: 1065-84.14 and van Wijngaarden, Coster and Williams (2005), JAMA, vol 293: 1509-13].
- Pan-VEGF inhibition appears to exert its beneficial effect mostly via an anti-permeability action resulting in resolution of intra- and sub-retinal edema, as the actual CNV lesion does not markedly involute [Presentation, at Angiogenesis 2006 Meeting. 2006. Bascom Palmer Eye Institute Miami, Florida].
- the lack of marked CNV involution may in part be a result of maturation of the newly formed vessels due to pericyte coverage.
- Pericytes play a critical role in the development and maintenance of vascular tissue.
- Remodeling of the vascular network involves adjustments in BV density to meet nutritional needs [Gariano and Gardner (2005), Nature, 438: 960-6]. Periods of BV immaturity corresponds to a period in which new vessels are functioning but have not yet acquired a pericyte coating [Benjamin, Hemo and Keshet (1998), Development, 125: 1591-8 and Gerhardt and Betsholtz (2003), Cell Tissue Res, 2003. 314: 15-23]. This delay is essential in providing a window of plasticity for the fine tuning of the developing vasculature according to the nutritional needs of the retina or choroid.
- the bioactive lipid sphingosine-1 -phosphate (SlP), VEGF, PDGF, angiopoietins (Ang) and other growth factors (GF) augment blood vessel growth and recruit smooth muscle cells (SMC) and pericytes to naive vessels which promote the remodeling of emerging vessels [Allende and Proia (2002), Biochim Biophys Acta, vol 582: 222-7; Gariano and Gardner (2005), Nature, vol 438: 960-6; Grosskreutz et al. (1999), Microvasc Res, vol 58: 128-36; Nishishita, and Lin (2004), J Cell Biochem, vol 91: 584-93 and Erber et al.
- SlP bioactive lipid sphingosine-1 -phosphate
- VEGF vascular endothelial growth factor
- PDGF angiopoietins
- Ang angiopoietins
- GF smooth muscle cells
- Pericytes most likely generated by in situ differentiation of mesenchymal precursors at the time of EC sprouting or from the migration and de-differentiation of arterial smooth muscle cells, intimately associate and ensheath ECs resulting in overall vascular maturity and survival [Benjamin, Hemo and Keshet (1998), Development, vol 125: 1591-8]. Recent studies have demonstrated that SlP, and the SlPl receptor, are involved in cell-surface trafficking and activation of the cell-cell adhesion molecule N-cadherin [Paik et al. (2004), Genes Dev, vol 18: 2392-403].
- N-cadherin is essential for interactions between EC, pericytes and mural cells which promote the development of a stable vascular bed [Gerhardt and Betsholtz (2003), Cell Tissue Res, vol 314: 15-23].
- Global deletion of the SlPl gene results in aberrant mural cell ensheathment of nascent BVs required for BV stabilization during embryonic development [Allende and Proia (2002), Biochim Biophys Acta, vol 1582: 222-7].
- Local injection of siRNA to SlPl suppresses vascular stabilization in tumor xenograft models [Chae et al. (2004), J Clin Invest, vol 114: 1082-9].
- VEGF vascular endothelial growth factor
- PDGF-B vascular endothelial growth factor-B promotes the maturation and stabilization of new BVs [Guo et al. (2003), Am J Pathol, 162: 1083-93 and Gariano and Gardner (2005), Nature, vol 438: 960-6.50].
- VEGF up-regulates Ang-1 (mRNA and protein) [Asahara et al. (1998), Circ Res, vol 83: 233-40].
- Ang-1 plays a major role in recruiting and sustaining peri-endothelial support by pericytes [Asahara et al. (1998), Circ Res, vol 83: 233-40].
- VEGF-B deficient mouse embryos lack micro-vascular pericytes, which leads to edema, micro-aneurisms and lethal hemorrhages [Lindahl et al. (1997), Science, vol 277: 242-5].
- Murine pre-natal studies have demonstrated that additional signals are required for complete VEGF- and PDGF-stimulation of vascular bed maturation. Based upon the trans-activation of SlP noted above, this factor could be SlP [Erber et al. (2004), FASEB J, vol 18: 338-40].
- Vessel stabilization and maturation is associated with a loss of plasticity and the absence of regression to VEGF and other GF withdrawal and resistance to anti-angiogenic therapies [Erber et al. (2004), FASEB J, vol 18: 338-40 and Hughes, and Chan-Ling (2004), Invest Ophthalmol Vis Sci, vol 45: 2795-806].
- Resistance of BVs to angiogenic inhibitors is conferred by pericytes that initially stabilize matured vessels and those that are recruited to immature vessels upon therapy [Erber et al. (2004), FASEB J, vol 18: 338-40]. After ensheathment of the immature ECs, the pericytes express compensatory survival factors (Ang-1 and PDGF-B) that protect ECs from pro-apoptotic agents.
- Ang-1 and PDGF-B compensatory survival factors
- CNV membranes are composed of fenestrated vascular ECs that tend to leak their intravascular contents into the surrounding space resulting in subretinal hemorrhage, exudates and fluid accumulation [Gerhardt and Betsholtz (2003), Cell Tissue Res, vol 14: 15-23].
- CNV tissue itself, and more recently intra-retinal neovascularization have been implicated as being responsible for the decrease in visual acuity associated with AMD.
- subretinal fibrosis leads to irreversible damage to the photoreceptors and permanent vision loss.
- the potential for subretinal fibrosis and future vision loss persists.
- RANIBIZUMAB it was discovered that those patients who lost vision did so as, a result of either subretinal fibrosis or a RPE tear [Presentation, at Angiogenesis 2006 Meeting. 2006. Bascom Palmer Eye Institute Miami, Florida.].
- An agent that could diminish the degree of fibroblast infiltration and collagen deposition would likely be of value.
- Fibroblasts are key cellular elements in scar formation in response to cellular injury and inflammation [Tomasek et al. (2002), Nat Rev MoI Cell Biol, vol 3: 349-63 and Virag and Murry (2003), Am J Pathol, vol 163: 2433-40]. Collagen gene expression by myofibroblasts is a hallmark of remodeling and necessary for scar formation [Sun and Weber (2000), Cardiovasc Res, vol 46: 250-6 and Sun and Weber (1996), J MoI Cell Cardiol, vol 28: 851-8]. SlP promotes wound healing by activating fibroblast migration and proliferation while increasing collagen production [Sun et al. (1994), J Biol Chem, vol 269: 16512-7].
- SlP produced locally by damaged cells could be responsible for the maladaptive wound healing associated with remodeling and scar formation.
- S IP inhibitors are useful in diseases or conditions characterized, at least in part, by aberrant fibrogenesis or fibrosis.
- fibrogenesis is defined as excessive activity or number of fibroblasts
- fibrosis is defined as excessive activity or number of fibroblasts that leads to excessive or inappropriate collagen production and scarring, destruction of the physiological tissue structure and/or inappropriate contraction of the matrix leading to such pathologies as retinal detachment or other processes leading to impairment of organ function.
- bioactive signaling lipids such as SlP and LPA is not limited to ocular diseases and conditions. Because of the involvement of biolipid signaling in many processes, including neovascularization, angiogenesis, aberrant fibrogenesis, fibrosis and scarring, and inflammation and immune responses, it is believed that antibody-based inhibitors of these bioactive lipids will be helpful in a variety of diseases and conditions associated with one or more of these processes. Such diseases and conditions may be systemic (e.g., systemic scleroderma) or localized to one or more specific body parts or organs (e.g., skin, lung, or eye).
- systemic e.g., systemic scleroderma
- specific body parts or organs e.g., skin, lung, or eye
- Lipids and their derivatives are now recognized as important targets for medical research, not as just simple structural elements in cell membranes or as a source of energy for ⁇ -oxidation, glycolysis or other metabolic processes.
- certain bioactive lipids function as signaling mediators important in animal and human disease.
- Most of the lipids of the plasma membrane play an exclusively structural role, a small proportion of them are involved in relaying extracellular stimuli into cells.
- “Lipid signaling” refers to any of a number of cellular signal transduction pathways that use cell membrane lipids as second messengers, as well as referring to direct interaction of a lipid signaling molecule with its own specific receptor.
- Lipid signaling pathways are activated by a variety of extracellular stimuli, ranging from growth factors to inflammatory cytokines, and regulate cell fate decisions such as apoptosis, differentiation and proliferation.
- Research into bioactive lipid signaling is an area of intense scientific investigation as more and more bioactive lipids are identified and their actions characterized.
- bioactive lipids include the eicosanoids (including the cannabinoids, leukotrienes, prostaglandins, lipoxins, epoxyeicosatrienoic acids, and isoeicosanoids), non- eicosanoid cannabinoid mediators, phospholipids and their derivatives such as phosphatidic acid (PA) and phosphatidylglycerol (PG), platelet activating factor (PAF) and cardiolipins as well as lysophospholipids such as lysophosphatidyl choline (LPC) and various lysophosphatidic acids (LPA).
- eicosanoids including the cannabinoids, leukotrienes, prostaglandins, lipoxins, epoxyeicosatrienoic acids, and isoeicosanoids
- non- eicosanoid cannabinoid mediators include phospholipids and their derivatives such as phosphatidic acid (PA) and
- Bioactive signaling lipid mediators also include the sphingolipids such as sphingomyelin, ceramide, ceramide-1-phosphate, sphingosine, sphingosylphosphoryl choline, sphinganine, sphinganine-1 -phosphate (Dihydro-S IP) and sphingosine- 1 -phosphate.
- Sphingolipids and their derivatives represent a group of extracellular and intracellular signaling molecules with pleiotropic effects on important cellular processes.
- bioactive signaling lipids include phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylethanolamine (PEA), diacylglyceride (DG), sulfatides, gangliosides, and cerebrosides.
- PS phosphatidylserine
- PI phosphatidylinositol
- PEA phosphatidylethanolamine
- DG diacylglyceride
- sulfatides gangliosides, and cerebrosides.
- Lysophospholipids also known as lysolipids, are low molecular weight (typically less than about 500 dalton) lipids that contain a single hydrocarbon backbone and a polar head group containing a phosphate group. Some lysolipids are bioactive signaling lipids. Two particular examples of medically important bioactive lysolipids are LPA (glycerol backbone) and SlP (sphingoid backbone). The structures of selected LPAs, SlP, and dihydro SlP are presented below.
- LPA is not a single molecular entity but a collection of endogenous structural variants with fatty acids of varied lengths and degrees of saturation (Fujiwara et al (2005), J Biol Chem, vol. 280: 35038-35050).
- the structural backbone of the LPAs is derived from glycerol-based phospholipids such as phosphatidylcholine (PC) or phosphatidic acid (PA).
- PC phosphatidylcholine
- PA phosphatidic acid
- SlP lysosphingolipids
- SlP dihydro SlP
- SPC sphingosylphosphorylcholine
- SPC sphingosylphosphorylcholine
- LPA and SlP regulate various cellular signaling pathways by binding to the same class of multiple transmembrane domain G protein-coupled (GPCR) receptors (Chun J, Rosen H (2006), Current Pharm Des, vol. 12: 161-171 and Moolenaar WH (1999), Experimental Cell Research, vol. 253: 230-238).
- the SlP receptors are designated as SlP 1 , SlP 2 , SlP 3 , SlP 4 and SlP 5 (formerly EDG-I, EDG-5/AGR16, EDG-3, EDG-6 and EDG-8) and the LPA receptors designated as LPA 1 , LPA 2 , LPA 3 (formerly, EDG-2, EDG-4, and EDG-7).
- a fourth LPA receptor of this family has been identified for LPA (LPA 4 ), and other putative receptors for these lysophospholipids have also been reported.
- S IP is a mediator of cell proliferation and protects from apoptosis through the activation of survival pathways (Maceyka et al. (2002), BBA, vol 1585): 192-201 and Spiegel S. et al. (2003), Nature Reviews Molecular Cell Biology, vol 4: 397-407). It has been proposed that the balance between ceramide/sphingosine (CER/SPH) levels and SlP provides a rheostat mechanism that decides whether a cell is directed into the death pathway or is protected from apoptosis.
- CER/SPH ceramide/sphingosine
- SlP The key regulatory enzyme of the rheostat mechanism is sphingosine kinase (SPHK) whose role is to convert the death-promoting bioactive signaling lipids (CER/SPH) into the growth-promoting SlP.
- SPHK sphingosine kinase
- CER/SPH death-promoting bioactive signaling lipids
- SlP has two fates: SlP can be degraded by SlP lyase, an enzyme that cleaves SlP to phosphoethanolamine and hexadecanal, or, less common, hydrolyzed by SlP phosphatase to SPH.
- S IP is abundantly generated and stored in platelets, which contain high levels of SPHK and lacks the enzymes for SlP degradation. When platelets are activated, SlP is secreted.
- SlP secreting protein
- carrier proteins such as serum albumin and lipoproteins.
- SlP is found in high concentrations in plasma, with concentrations in the range of 0.5 - 5 uM having been reported.
- intracellular actions of SlP have also been suggested (see, eg, Spiegel S, Kolesnick R (2002), Leukemia, vol. 16: 1596-602; Suomalainen, et al (2005), Am J Pathol, vol. 166: 773- 81).
- SlP receptors Widespread expression of the cell surface SlP receptors allows SlP to influence a diverse spectrum of cellular responses, including proliferation, adhesion, contraction, motility, morphogenesis, differentiation, and survival. This spectrum of response appears to depend upon the overlapping or distinct expression patterns of the S IP receptors within the cell and tissue systems.
- crosstalk between SlP and growth factor signaling pathways including platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), transforming growth factor beta (TGF ⁇ ) and basic fibroblastic growth factor (bFGF), have recently been demonstrated (see, e.g., Baudhuin, et al (2004), FASEB J, vol. 18: 341-3).
- SPHINGOMAB murine monoclonal antibody
- SPHINGOMAB represents the first successfully created monoclonal antibody against a bioactive signaling sphingolipid target.
- SPHINGOMAB acts as a molecular sponge to selectively absorb SlP from the extracellular fluid, lowering the effective concentration of SlP. It selectively binds and neutralizes SlP with picomolar affinity in biologic matrices.
- SPHINGOMAB would deprive fibroblasts, pericytes, and endothelial, inflammatory and immune cells in the eye of important growth and survival factors thus targeting the multiple maladaptive steps of AMD resulting in the loss of photoreceptors and visual acuity.
- a therapeutic that simultaneously targets multiple components of the choroidal neovascular response has the potential to be a more potent therapeutic than "single-target" therapeutics.
- sphingosine-l-phosphate or “SlP” refers to sphingosine-1 -phosphate [sphingene-1 -phosphate; D-erythro-sphingosine-1-phosphate; sphing-4-enine-l -phosphate; (E,2S,3R)-2-amino-3-hydroxy-octadec-4-enoxy]phosphonic acid; CAS
- SlP and DHSlP dihydro sphingosine-1 -phosphate [sphinganine-1-phosphate; [(2S,3R)-2-amino-3-hydroxy-octadecoxy]phosphonic acid; D- Erythro-dihydro-D-sphingosine-l-phosphate;CAS 19794-97-9] and sphingosylphosphorylcholine.
- "Variants" of SlP and LPA includes analogs and derivatives of SlP and LPA, respectively, which function similarly, or might be expected to function similarly, to the parent molecule.
- SlP could contribute to both the early and late stages of maladaptive retinal remodeling associated with exudative AMD.
- SlP has a pronounced non- VEGF dependent pro-angiogenic effect.
- SlP also stimulates migration, proliferation and survival of multiple cell types, including fibroblasts, EC, pericytes and inflammatory cells — the same cells that participate in the multiple maladaptive processes of exudative AMD.
- SlP is linked to the production and activation of VEGF, bFGF, PDGF and other growth factors (GFs) implicated in the pathogenesis of exudative AMD.
- SlP may modulate the maturation of na ⁇ ve vasculature, a process leading to a loss of sensitivity to anti-angiogenic agents.
- Inhibiting the action of S IP could be an effective therapeutic treatment for exudative AMD that may offer significant advantages over exclusively anti-VEGF approaches or may act synergistically with them to address the complex processes and multiple steps that ultimately lead to AMD associated visual loss.
- SlP is an important mediator of inflammatory events [Olivera and Rivera (2005), J Immunol,, vol 174: 1153-8]. Activated platelets, neutrophils, macrophages and mast cells serve as rich sources of SlP after coagulation and inflammatory events [Yatomi et al. (2000) Blood, vol 96: 3431-8]. Because these cells are important components in the inflammation response and tissue loss, SlP may regulate these events via control of inflammatory cell function [Tezel (2004), Trends MoI Med, vol 10: 417-20]. SlP released from mast cells is responsible for many of the responses in experimental animal models of inflammation [Jolly et al.
- Neutralizing SlP with SPHINGOMAB could provide an effective, novel means of limiting the deleterious inflammatory response that exacerbates ocular tissue damage of CNV associated with AMD.
- SlP stimulates the formation and maintenance of vascular EC assembly and integrity by activating both SlP 1 and SlP 3 , and S IP-induced EC adherent junction assembly [Paik et al. (2004), Genes Dev, vol 18: 2392-403 and Lee et al. (1999), Cell, vol 99: 301-12].
- Antisense oligonucleotides against these SlP receptors diminish SlP-induced vascular EC assembly and cell barrier integrity [English, et al.
- SlP elicits a synergic effect with VEGF, EGF, PDGF, bFGF and IL-8 to promote the development of vascular networks in vivo [Wang et al. (1999), J Biol. Chem., vol 274: 35343-50].
- SlP trans-activates EGF and VEGF2 receptors [Tanimoto, Jin and Berk (2002), J Biol Chem, vol 277: 42997-3001] and VEGF up-regulates SlP receptors [Igarashi et al. (2003), Proc Natl Acad Sci U S A, vol 100: 10664-9].
- vascular ECs Treatment of vascular ECs with VEGF markedly induces the up-regulation of SlPl expression and enhances S IP-mediated signaling pathways leading to the activation of the endothelial isoform of nitric oxide synthase (eNOS) [Lee et al (2001), MoI Cell, vol 8: 693-704 and Tanimoto, Jin and Berk (2002), J Biol Chem, vol 277: 42997-3001 and Igarashi and Michel (2001), J Biol Chem, vol 276: 36281-8].
- eNOS activity plays a crucial role in different cellular responses and essential vascular functions, including inhibition of apoptosis, inhibition of platelet aggregation and angiogenesis [Kwon et al.
- SPHINGOMAB may mitigate aberrant BV growth by neutralizing synergistic pro- angiogenic GFs and possibly SlP produced in excess during metabolic stress from inflammatory cells associated with CNV.
- SPHINGOMAB not only inhibits S IP-induced EC migration/infiltration and BV formation, but it also neutralizes bFGF and VEGF-induced vascularization through its effect on SlP.
- SPHINGOMAB has a potential advantage over "single-target" therapeutics because of its ability to neutralize SlP, which results in neutralization of multiple GFs via the pleiotropic effects of SlP.
- Direct neutralization of SlP and an indirect neutralization of VEGF and PDGF-B by SPHINGOMAB could prevent pericyte recruitment, BV maturation and slow the development of resistance to anti-angiogenic drugs.
- Targeting pericytes, in the effort to extended or increase vulnerability to anti-angiogenic agents, represents an attractive long-term approach in treating patients presenting with active CNV lesions and could promote involution of vascular complexes [Erber et al. (2004), FASEB J, vol 18: 338-40].
- VEGF and PDGF can compromise blood-retinal barrier (BRB) integrity: SPHINGOMAB' s ability to neutralize SlP trans-activation of VEGF and PDGF could prove effective in mitigating macular edema associated with AMD [Sanchez et al. (2003), J Biol Chem, vol 278: 47281-90; Saishin et al (2003), J Cell Physiol, vol 195: 241-8 and Vinores et al (2000), Gen Pharmacol, vol 35: 233-9].
- Transgenic mice overexpressing VEGF demonstrate a BRB breakdown occurring in the area of CNV similar to that seen in AMD and diabetic retinopathies [Vinores et al.
- Fibroblasts respond to SlP treatment by an increase in DNA synthesis; fibroblasts transfected with Sphingosine Kinase 1 (sphKl) exhibit increased cellular proliferation [Hammer et al (2004), J Cell Biochem, vol 91: 840-51]. Similar to the effects of SlP on several other fibroblast types (Swiss 3T3, lung and cardiac), SlP may stimulate ocular fibroblast proliferation (and subsequent differentiation).
- Fibroblasts are directly protected from apoptosis by addition of SlP, and apoptosis is enhanced by inhibitors of sphKl [Olivera et al (1999), J Cell Biol, vol 147: 545-58].
- SlP blocks cytochrome C release and subsequent caspase activation [Olivera et al (1999), J Cell Biol, vol 147: 545-58 and Kang et al. (2004), Cell Death Differ, vol 11: 1287-98]. It is established that sphKl upregulates Akt, thereby regulating Bcl-2 family members [Limaye et al. (2005), Blood, vol 105: 3169-77] and protecting fibroblasts from apoptosis.
- SlP and fibroblast migration SlP activates signaling systems including Rho, resulting in the assembly of contractile actin filaments controlled by Rho/Rac/Cdc42 system, and leading to substantial effects on cellular migration [Radeff-Huang et al. (2004), J Cell Biochem,. Vol 92: 949-66].
- Rho and Rho GTPases by SlP may be responsible for the migration of ocular fibroblasts into the wound and thereby contribute to fibrosis.
- SlP and fibroblast collagen expression SlP promotes the differentiation of quiescent fibroblasts to active myofibroblasts which exhibit enhanced collagen expression during scar formation [Urata et al. (2005), Kobe J Med Sci, vol 51: 17-27]. Concurrent with the proliferation and migration of fibroblasts into the scarring zone, myofibroblasts deposit a temporary granular network consisting primarily of osteopontin and fibronectin [Sun and Weber (2000), Cardiovasc Res, vol 46: 250-6]. As remodeling proceeds, the temporary matrix is absorbed and a collagen network established [Sun and Weber (2000), Cardiovasc Res, vol 46: 250-6]. We have demonstrated that SlP promotes collagen production by myofibroblasts.
- TGF ⁇ a well-known fibrotic mediator
- TGF ⁇ a well-known fibrotic mediator
- SPKQNGOMAB could mitigate the profibrotic effects of TGF ⁇ as well as mitigating the fibrogenic effects of SlP itself.
- Minimizing maladaptive scar formation by neutralization of SlP could be beneficial and prevent irreversible losses in visual acuity by limiting the extent of sub- retinal fibrosis and subsequent photoreceptor damage.
- LPA Lysophosphatic acids
- LPA have long been known as precursors of phospholipid biosynthesis in both eukaryotic and prokaryotic cells, but LPA have emerged only recently as signaling molecules that are rapidly produced and released by activated cells, notably platelets, to influence target cells by acting on specific cell-surface receptor (see, eg, Moolenaar et al. (2004), BioEssays, vol. 26: 870-881 and van Leewen et al. (2003), Biochem Soc Trans, vol 31: 1209-1212).
- LPA can be generated through the hydrolysis of pre-existing phospholipids following cell activation; for example, the sn-2 position is commonly missing a fatty acid residue due to de-acylation, leaving only the sn-3 hydroxyl esterified to a fatty acid.
- autotaxin lysoPLD/NPP2
- lysoPLD/NPP2 may be the product of an oncogene, as many tumor types up-regulate autotaxin (Brindley (2004), J Cell Biochem, vol. 92: 900-12).
- LPA concentrations in human plasma and serum have been reported, including determinations made using sensitive and specific LC/MS procedures (Baker et al. (2001), Anal Biochem, vol 292: 287-295).
- LPA concentrations have been estimated to be approximately 1.2 ⁇ M, with the LPA analogs 16:0, 18:1, 18:2, and 20:4 being the predominant species.
- LPA concentrations have been estimated to be approximately 0.7 ⁇ M, with 18:1 and 18:2 LPA being the predominant species.
- LPA influence a wide range of biological responses, including induction of cell proliferation, stimulation of cell migration and neurite retraction, gap junction closure, and even slime mold chemotaxis (Goetzl. et al. (2002), Scientific World Journal, vol 2: 324-338).
- the body of knowledge about the biology of LPA continues to grow as more and more cellular systems are tested for LPA responsiveness. For instance, it is now known that, in addition to stimulating cell growth and proliferation, LPA promote cellular tension and cell-surface fibronectin binding, which are important events in wound repair and regeneration (Moolenaar et al. (2004), BioEssays, vol. 26: 870-881).
- peroxisome proliferation receptor gamma is a receptor/target for LPA (Simon et al. (2005), J Biol Chem, vol 280: 14656-14662).
- the anti-LPA antibodies can neutralize various LPAs and mitigate their biologic and pharmacologic action.
- the anti-LPA antibodies would be expected to act on the following processes for therapeutic benefit.
- LPA Edema and vascular permeability: LPA induces plasma exudation and histamine release in mice [Hashimoto et al (2006), J Pharmacol Sci, vol 100: 82-7].
- LPA acts as inflammatory mediator in human corneal epithelial cells [Zhang et al (2006), Am J Physiol, June 7]. LPA participates in corneal wound healing [Liliom K et al (1998), Am. J. Physiol, vol 274: C1065-C1074] and stimulates the release of ROS in lens tissue [Rao et al. (2004), Molecular Visions, vol 10: 112-121]. LPA can also re-activate HSV-I in rabbit cornea [Martin et al. (1999), Molecular Visions, vol 5: 36-42 ⁇ .
- LPA inhibits TGF ⁇ -mediated stimulation of type I collagen mRNA stability via an ERK-dependent pathway in dermal fibroblasts [Sato et al (2004), Matrix Biol, vol 23: 353-61]. Moreover, LPA have some direct fibrogenic effects by stimulating collagen gene expression and proliferation of fibroblasts [ Chen, et al. (2006) FEBS Lett. 580(19):4737-45.
- immuno-derived moiety refers to any polyclonal or monoclonal antibody or antibody fragment, variant, or derivative.
- an “anti-SIP antibody” or an “immune-derived moiety reactive against SlP” refers to any antibody or antibody-derived molecule that binds SlP.
- an “anti-LPA antibody” or an “immune-derived moiety reactive against LPA” refers to any antibody or antibody-derived molecule that binds to all or one or more of the LPAs.
- a “bioactive lipid” refers to a lipid signaling molecule.
- a bioactive lipid does not reside in a biological membrane when it exerts its signaling effects, which is to say that while such a lipid species may exist at some point in a biological membrane (for example, a cell membrane, a membrane of a cell organelle, etc.), when associated with a biological membrane it is not a “bioactive lipid” but is instead a "structural lipid” molecule.
- Bioactive lipids are distinguished from structural lipids (e.g., membrane-bound phospholipids) in that they mediate extracellular and/or intracellular signaling and thus are involved in controlling the function of many types of cells by modulating differentiation, migration, proliferation, secretion, survival, and other processes.
- structural lipids e.g., membrane-bound phospholipids
- bioactive lipids can be found in extracellular fluids, where they can be complexed with other molecules, for example serum proteins such as albumin and lipoproteins, or in "free" form, i.e., not complexed with another molecule species.
- bioactive lipids alter cell signaling by activating membrane-bound ion channels or G-protein coupled receptors that, in turn, activate complex signaling systems that result in changes in cell function or survival.
- bioactive lipids can exert their actions by directly interacting with intracellular components such as enzymes and ion channels.
- Representative examples of bioactive lipids include LPA and SlP.
- the term "therapeutic agent” means an agent to mitigate angiogenesis and/or neovascularization, e.g., CNV and BV maturation; edema, vascular permeability and fibrosis, fibrogenesis and scarring associated with, or part of the underlying pathology of, ocular diseases and conditions.
- combination therapy refers to a therapeutic regimen that involves the provision of at least two distinct therapies to achieve an indicated therapeutic effect.
- a combination therapy may involve the administration of two or more chemically distinct active ingredients, for example, an anti-LPA antibody and an anti-SIP antibody.
- a combination therapy may involve the administration of an immune-derived moiety reactive against a bioactive lipid and the administration of one or more other chemotherapeutic agents.
- Combination therapy may, alternatively, involve administration of an anti-lipid antibody together with the delivery of another treatment, such as radiation therapy and/or surgery.
- a combination therapy may involve administration of an anti-lipid antibody together with one or more other biological agents (e.g., anti- VEGF, TGF ⁇ , PDGF, or bFGF agent), chemotherapeutic agents and another treatment such as radiation and/or surgery.
- the active ingredients may be administered as part of the same composition or as different compositions.
- the compositions comprising the different active ingredients may be administered at the same or different times, by the same or different routes, using the same of different dosing regimens, all as the particular context requires and as determined by the attending physician.
- the drug(s) may be delivered before or after surgery or radiation treatment.
- “Monotherapy” refers to a treatment regimen based on the delivery of one therapeutically effective compound, whether administered as a single dose or several doses over time.
- a "patentable" composition, process, machine, or article of manufacture according to the invention means that the subject matter satisfies all statutory requirements for patentability at the time the analysis is performed. For example, with regard to novelty, non-obviousness, or the like, if later investigation reveals that one or more claims encompass one or more embodiments that would negate novelty, non-obviousness, etc., the claim(s), being limited by definition to “patentable” embodiments, specifically exclude the unpatentable embodiment(s). Also, the claims appended hereto are to be interpreted both to provide the broadest reasonable scope, as well as to preserve their validity.
- pharmaceutically acceptable salt refers to salts which retain the biological effectiveness and properties of the agents and compounds of this invention and which are not biologically or otherwise undesirable.
- the agents and compounds of this invention are capable of forming acid and/or base salts by virtue of the presence of charged groups, for example, charged amino and/or carboxyl groups or groups similar thereto.
- Pharmaceutically acceptable acid addition salts may be prepared from inorganic and organic acids, while pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases.
- sample-holding vessel The terms “separated,” “purified,” “isolated,” and the like mean that one or more components of a sample contained in a sample-holding vessel are or have been physically removed from, or diluted in the presence of, one or more other sample components present in the vessel.
- Sample components that may be removed or diluted during a separating or purifying step include, chemical reaction products, unreacted chemicals, proteins, carbohydrates, lipids, and unbound molecules.
- kits is used herein in various contexts, e.g., a particular species of chemotherapeutic agent.
- the term refers to a population of molecules, chemically indistinguishable from each other, of the sort referred in the particular context.
- Specifically associate and “specific association” and the like refer to a specific, non- random interaction between two molecules, which interaction depends on the presence of structural, hydrophobic/hydrophilic, and/or electrostatic features that allow appropriate chemical or molecular interactions between the molecules.
- stable refers to an interaction between two molecules (eg, binding of an anti- LPA or anti-SlP antibody to its target bioactive lipid) that is sufficiently strong such that the molecules can be maintained for the desired purpose or manipulation.
- a “subject” or “patient” refers to an animal in which treatment can be effected by molecules of the invention.
- the animal may have, be at risk for, or be believed to have or be at risk for a disease or condition that can be treated by compositions and/or methods of the present invention.
- Animals that can be treated in accordance with the invention include vertebrates, with mammals such as bovine, canine, equine, feline, ovine, porcine, and primate (including humans and non-human primates) animals being particularly preferred examples.
- a “therapeutically effective amount” refers to an amount of an active ingredient, e.g., an agent according to the invention, sufficient to effect treatment when administered to a subject or patient. Accordingly, what constitutes a therapeutically effective amount of a composition according to the invention may be readily determined by one of ordinary skill in the art.
- a “therapeutically effective amount” is one that produces an objectively measured change in one or more parameters associated with treatment of the ocular disease or condition including an increase or decrease in the expression of one or more genes correlated with the ocular disease or condition, induction of apoptosis or other cell death pathways, clinical improvement in symptoms, a decrease in aberrant neovascularization or in inflammation, etc.
- the therapeutically effective amount will vary depending upon the particular subject and condition being treated, the weight and age of the subject, the severity of the disease condition, the particular compound chosen, the dosing regimen to be followed, timing of administration, the manner of administration and the like, all of which can readily be determined by one of ordinary skill in the art. It will be appreciated that in the context of combination therapy, what constitutes a therapeutically effective amount of a particular active ingredient may differ from what constitutes a therapeutically effective amount of the active ingredient when administered as a monotherapy (ie. , a therapeutic regimen that employs only one chemical entity as the active ingredient).
- treatment or “treating” of a disease or disorder includes preventing or protecting against the disease or disorder (that is, causing the clinical symptoms not to develop); inhibiting the disease or disorder (i.e., arresting or suppressing the development of clinical symptoms; and/or relieving the disease or disorder (i.e., causing the regression of clinical symptoms).
- preventing and “suppressing” a disease or disorder since the ultimate inductive event or events may be unknown or latent.
- the term “prophylaxis” will be understood to constitute a type of “treatment” that encompasses both "preventing” and “suppressing.” The term “treatment” thus includes “prophylaxis”.
- therapeutic regimen means any treatment of a disease or disorder using chemotherapeutic drugs, radiation therapy, surgery, gene therapy, DNA vaccines and therapy, antisense-based therapies including siRNA therapy, anti-angiogenic therapy, immunotherapy, bone marrow transplants, aptamers and other biologies such as antibodies and antibody variants, receptor decoys and other protein-based therapeutics.
- a pharmaceutical composition comprising an immune-derived moiety (e.g, an antibody) reactive against a bioactive lipid, in order to decrease the effective concentration so that the bioactive lipid is inhibited in whole or in part from eliciting its undesired effects.
- an immune-derived moiety e.g, an antibody
- the immune-derived moiety is a monoclonal antibody or fragment, variant or derivative thereof.
- the immune-derived moiety is reactive against a lysolipid, such as SlP or LPA.
- Methods are also provided for decreasing or preventing aberrant fibrogenesis, fibrosis or scarring; inflammation; or aberrant neovascularization; modulating surgical and traumatic wound healing responses of the eye; or for attenuating an ocular immune response. Further provided are methods for decreasing the effective ocular concentration or activity of bioactive lipid. Also provided are methods of treating scleroderma using an immune-derived moiety reactive against a bioactive lipid, such as the lysolipids SlP or LPA.
- Representative bioactive lipids include sphingolipids and variants thereof such as sphingosine-1 -phosphate (SlP), sphingosine, sphingosylphosphorylcholine, dihydrosphingosine.
- Other bioactive lysolipids include lysophosphatidic acids (LPAs) and variants thereof.
- compositions including those for ocular administration, that comprise a carrier and an isolated immune-derived moiety, for example, a monoclonal antibody or antibody fragment, variant, or derivative, reactive against a bioactive lipid.
- Preferred carriers include those that are pharmaceutically acceptable, particularly when the composition is intended for therapeutic use in humans.
- veterinarily acceptable carriers may be employed.
- Exemplary routes of administration of an immune-derived moiety according to the invention include systemic administration, parenteral administration (e.g., via injection via an intravenous, intramuscular, or subcutaneous route), transdermal, intradermal or transmucosal delivery, intraocular or periocular injection, mucosal or topical administration or by inhalation.
- parenteral administration e.g., via injection via an intravenous, intramuscular, or subcutaneous route
- transdermal e.g., intradermal or transmucosal delivery
- intraocular or periocular injection e.g., mucosal or topical administration or by inhalation.
- FIG. 1 SPHINGOMAB reduced CNV and scar formation in ocular lesions. Mice were treated with SPHINGOMAB or an isotype-matched non-specific mAb. CNV lesions were induced by laser rupture of Bruchs membrane. Shown are graphs and representative images of lesions from each treatment group stained with rhodamine-conjugated R. communis agglutinin I for vascularization (A) or Masson's Trichrome for collagen scar formation (B).
- Figure Ia shows that SPHINGOMAB dramatically attenuates choroidal neovascularization 14 and 28 days after laser-induced rupture of Bruch's membrane.
- Figure Ib shows that SPHINGOMAB significantly reduces fibrosis associated with CNV lesion formation 28 days after laser-induced rupture of Bruchs' s membrane.
- FIG. 2 SlP promotes neovascularization through induction of HUVECs tube formation and migration and is reduced by SPHINGOMAB.
- Panel A Micrographs of HUVECs seeded on Matrigel and incubated for 6 hrs to evaluate tube formation.
- Panel B HUVECs were treated with l ⁇ M SlP ⁇ SPHINGOMAB (l ⁇ g/ml) for 6 hrs in a Matrigel invasion chamber. The number of cells that migrated to the Matrigel membrane were counted in 5 independent fields.
- SPHINGOMAB neutralizes SlP-, VEGF- and bFGF-induced neovascularization.
- A Representative FITC-stained BVs from sections of Matrigel plugs ⁇ GFs.
- B SlP stimulates EC infiltration.
- C Quantification of relative fluorescence from Matrigel plugs stimulated with VEGF or bFGF as an indicator of neovascularization. SlP, VEGF and bFGF's effects were inhibited when mice were systemically treated with 1 or 25mg/kg of SPHINGOMAB.
- FIG. 4 SPHINGOMAB neutralized S IP-stimulated scar formation. Fibroblasts were serum-starved and then treated with 0, 0.1, 0.5 or 1/xM SlP +/- l ⁇ g/mL SPHINGOMAB for 12- 24 hrs. SlP stimulated Swiss 3T3 fibroblast proliferation as measured by 3H-thymidine incorporation (A), murine cardiac fibroblast migration in a scratch assay (B), collagen gene expression (relative fluorescence) in isolated cardiac fibroblasts from transgenic mice expressing collagen-GFP (C) and WI-38 cell differentiation into myofibroblasts as measured by decreased cellular proliferation and increased ⁇ -SMA expression (D); SPHINGOMAB neutralized each of S IP' s effects. SPHINGOMAB reduced perivascular fibrosis in vivo in a murine model of a permanent myocardial infarction (E).
- E myocardial infarction
- SlP promotes transformation of ocular epithelial cells and fibroblasts into contractile, scar tissue-producing myofibroblasts.
- the effects of SlP on myofibroblast transformation of several human ocular cell lines were examined.
- SlP was found to stimulate production of ⁇ -Smooth muscle actin ( ⁇ -SMA; a myofibroblast marker) in human retinal pigmented epithelial cells (Figure 5A) and human conjunctiva fibroblasts ( Figure 5B).
- ⁇ -SMA smooth muscle actin
- Figure 5A human retinal pigmented epithelial cells
- Figure 5B human conjunctiva fibroblasts
- PAI-I plasminogen activator inhibitor
- SPHINGOMAB reduced immune-cell wound infiltration in vivo. Mice were subjected to MI, treated with saline or 25mg/kg SPHINGOMAB 48 hrs after surgery and then sacrificed on day 4. SPHINGOMAB reduced macrophage (A) and mast cell (B) infiltration into the wound. Data are represented as fold decrease of saline treated values.
- SPHINGOMAB is highly specific for SlP.
- a graph based on competitive ELISA demonstrates SPHINGOMAB 's specificity for SlP compared to other bioactive lipids.
- SPHINGOMAB demonstrated no cross-reactivity to sphingosine (SPH), the immediate metabolic precursor of SlP or lysophosphatidic acid (LPA), an important extracellular signaling molecule that is structurally and functionally similar to SlP.
- SPH sphingosine
- LPA lysophosphatidic acid
- SPHINGOMAB did not recognize other structurally similar lipids and metabolites, including ceramide-1 -phosphate (ClP), dihydrosphingosine (DH-SPH), phosphatidyl serine (PS), phosphatidyl ethanolamine (PE), or sphingomyelin (SM). SPHINGOMAB did cross react with dihydrosphingosine- 1 -phosphate (DH-SlP) and, to a lesser extent, sphingosylphoryl choline (SPC). The affinity (Kd) of SPHINGOMAB for SlP is ⁇ 100pM, much higher than most therapeutic antibodies, particularly other molecular sponges.
- immunoglobulins refers to any form of a peptide, polypeptide derived from, modeled after or encoded by, an immunoglobulin gene, or a fragment of such peptide or polypeptide that is capable of binding an antigen or epitope [see, eg, Immunobiology, 5th Edition, Janeway, Travers, Walport, Shlomchiked. (editors), Garland Publishing (2001)].
- the antigen is a bioactive lipid molecule.
- Antibody molecules or immunoglobulins are large glycoprotein molecules with a molecular weight of approximately 150 kDa, usually composed of two different kinds of polypeptide chain.
- Each immunoglobulin molecule usually consists of two heavy chains and two light chains.
- the two heavy chains are linked to each other by disulfide bonds, the number of which varies between the heavy chains of different immunoglobulin isotypes.
- Each light chain is linked to a heavy chain by one covalent disulfide bond.
- the two heavy chains and the two light chains are identical, harboring two identical antigen- binding sites, and are thus said to be divalent, i.e., having the capacity to bind simultaneously to two identical molecules.
- the "light” chains of antibody molecules from any vertebrate species can be assigned to one of two clearly distinct types, kappa (k) and lambda (1), based on the amino acid sequences of their constant domains.
- the ratio of the two types of light chain varies from species to species. As a way of example, the average k to 1 ratio is 20:1 in mice, whereas in humans it is 2:1 and in cattle it is 1:20.
- the "heavy" chains of antibody molecules from any vertebrate species can be assigned to one of five clearly distinct types, called isotypes, based on the amino acid sequences of their constant domains. Some isotypes have several subtypes.
- the five major classes of immunoglobulin are immunoglobulin M (IgM), immunoglobulin D (IgD), immunoglobulin G (IgG), immunoglobulin A (IgA), and immunoglobulin E (IgE).
- IgG is the most abundant isotype and has several subclasses (IgGl, 2, 3, and 4 in humans).
- the Fc fragment and hinge regions differ in antibodies of different isotypes, thus determining their functional properties. However, the overall organization of the domains is similar in all isotypes.
- variable region refers to the N-terminal portion of the antibody molecule or a fragment thereof.
- each of the four chains has a variable (V) region in its amino terminal portion, which contributes to the antigen-binding site, and a constant (C) region, which determines the isotype.
- the light chains are bound to the heavy chains by many noncovalent interactions and by disulfide bonds and the V regions of the heavy and light chains pair in each arm of antibody molecule to generate two identical antigen-binding sites. Some amino acid residues are believed to form an interface between the light- and heavy-chain variable domains [see Kabat et al. (1991), Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md. and Clothia et al. (1985), J. MoI. Biol, vol 186: 651].
- variable domains of antibodies variability is not uniformly distributed throughout the variable domains of antibodies, but is concentrated in three segments called “complementarity-determining regions” (CDRs) or “hypervariable regions” both in the light-chain and the heavy-chain variable domains.
- CDRs complementarity-determining regions
- FR framework region
- the variable domains of native heavy and light chains each comprise four FR regions connected by three CDRs.
- the CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chains, form the antigen-binding site of antibodies [see Kabat et al. (1991), Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md.].
- constant domain refers to the C-terminal region of an antibody heavy or light chain.
- the constant domains are not directly involved in the binding properties of an antibody molecule to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular toxicity.
- effector functions refer to the different physiological effects of antibodies (e.g., opsonization, cell lysis, mast cell, basophil and eosinophil degranulation, and other processes) mediated by the recruitment of immune cells by the molecular interaction between the Fc domain and proteins of the immune system.
- the isotype of the heavy chain determines the functional properties of the antibody. Their distinctive functional properties are conferred by the carboxy-terminal portions of the heavy chains, where they are not associated with light chains.
- antibody fragment refers to a portion of an intact antibody that includes the antigen binding site or variable regions of an intact antibody, wherein the portion can be free of the constant heavy chain domains (e.g., CH2, CH3, and CH4) of the Fc region of the intact antibody.
- portions of the constant heavy chain domains e.g., CH2, CH3, and CH4 can be included in the "antibody fragment".
- antibody fragments are those that retain antigen-binding and include Fab, Fab', F(ab')2, Fd, and Fv fragments; diabodies; triabodies; single-chain antibody molecules (sc-Fv); minibodies, nanobodies, and multispecific antibodies formed from antibody fragments.
- a Fab fragment also contains the constant domain of a light chain and the first constant domain (CHl) of a heavy chain.
- variable refers to an amino acid sequence which differs from the native amino acid sequence of an antibody by at least one amino acid residue or modification.
- a native or parent or wild-type amino acid sequence refers to the amino acid sequence of an antibody found in nature.
- Variant of the antibody molecule includes, but is not limited to, changes within a variable region or a constant region of a light chain and/or a heavy chain, including the hypervariable or CDR region, the Fc region, the Fab region, the CH 1 domain, the CH 2 domain, the CH 3 domain, and the hinge region.
- the term "specific” refers to the selective binding of an antibody to its target epitope.
- Antibody molecules can be tested for specificity of binding by comparing binding of the antibody to the desired antigen to binding of the antibody to unrelated antigen or analogue antigen or antigen mixture under a given set of conditions.
- an antibody according to the invention will lack significant binding to unrelated antigens, or even analogs of the target antigen.
- the term "antigen” refers to a molecule that is recognized and bound by an antibody molecule or immune-derived moiety that binds to the antigen.
- a “hapten” refers to a small molecule that can, under most circumstances, elicit an immune response (i.e., act as an antigen) only when attached to a carrier molecule, for example, a protein, polyethylene glycol (PEG), colloidal gold, silicone beads, and the like.
- the carrier may be one that also does not elicit an immune response by itself.
- antibody is used in the broadest sense, and encompasses monoclonal, polyclonal, multispecific (e.g., bispecific, wherein each arm of the antibody is reactive with a different epitope or the same or different antigen), minibody, heteroconjugate, diabody, triabody, chimeric, and synthetic antibodies, as well as antibody fragments that specifically bind an antigen with a desired binding property and/or biological activity.
- mAb refers to an antibody, or population of like antibodies, obtained from a population of substantially homogeneous antibodies, and is not to be construed as requiring production of the antibody by any particular method.
- monoclonal antibodies can be made by the hybridoma method first described by Kohler and Milstein (1975), Nature, vol 256: 495-497, or by recombinant DNA methods.
- chimeric antibody refers to a molecule comprising a heavy and/or light chain which is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity [Cabilly et al. (1984), infra; Morrison et al, Proc. Natl. Acad. Sci. U.S.A. 81:6851].
- humanized antibody refers to forms of antibodies that contain sequences from non-human (eg, murine) antibodies as well as human antibodies.
- a humanized antibody can include conservative amino acid substitutions or non-natural residues from the same or different species that do not significantly alter its binding and/or biologic activity.
- Such antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulins.
- humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a complementary-determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat, camel, bovine, goat, or rabbit having the desired properties.
- CDR complementary-determining region
- humanized antibodies can comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences. These modifications are made to further refine and maximize antibody performance.
- a humanized antibody will comprise all of at least one, and in one aspect two, variable domains, in which all or all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence.
- the humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), or that of a human immunoglobulin. See, e.g., Cabilly et al, U.S. Pat. No.
- bispecific antibody can refer to an antibody, or a monoclonal antibody, having binding properties for at least two different epitopes.
- the epitopes are from the same antigen.
- the epitopes are from two different antigens.
- Methods for making bispecific antibodies are known in the art. For example, bispecific antibodies can be produced recombinantly using the co-expression of two immunoglobulin heavy chain/light chain pairs. Alternatively, bispecific antibodies can be prepared using chemical linkage. Bispecific antibodies include bispecific antibody fragments.
- heteroconjugate antibody can refer to two covalently joined antibodies. Such antibodies can be prepared using known methods in synthetic protein chemistry, including using crosslinking agents. As used herein, the term “conjugate” refers to molecules formed by the covalent attachment of one or more antibody fragment(s) or binding moieties to one or more polymer molecule(s).
- biologically active refers to an antibody or antibody fragment that is capable of binding the desired epitope and in some way exerting a biologic effect.
- Biological effects include, but are not limited to, the modulation of a growth signal, the modulation of an anti- apoptotic signal, the modulation of an apoptotic signal, the modulation of the effector function cascade, and modulation of other ligand interactions.
- recombinant DNA refers to nucleic acids and gene products expressed therefrom that have been engineered, created, or modified by man.
- Recombinant polypeptides or proteins are polypeptides or proteins produced by recombinant DNA techniques, for example, from cells transformed by an exogenous DNA construct encoding the desired polypeptide or protein.
- Synthetic polypeptides or proteins are those prepared by chemical synthesis.
- expression cassette refers to a nucleotide molecule capable of affecting expression of a structural gene (i.e., a protein coding sequence, such as an antibody of the invention) in a host compatible with such sequences.
- Expression cassettes include at least a promoter operably linked with the polypeptide-coding sequence, and, optionally, with other sequences, e.g., transcription termination signals. Additional regulatory elements necessary or helpful in effecting expression may also be used, e.g., enhancers.
- expression cassettes include plasmids, expression vectors, recombinant viruses, any form of recombinant "naked DNA" vector, and the like.
- the invention is drawn to compositions and methods for treating or preventing ocular diseases and conditions, using one or more therapeutic agents that alter the activity or concentration of one or more undesired bioactive lipids, or precursors or metabolites thereof.
- the therapeutic methods and compositions of the invention act by changing the effective concentration, i.e., the absolute, relative, effective and/or available concentration and/or activities, of certain undesired bioactive lipids. Lowering the effective concentration of the bioactive lipid may be said to "neutralize" the target lipid or its undesired effects, including downstream effects.
- “undesired” refers to a bioactive lipid that is unwanted due to its involvement in a disease process, for example, as a signaling molecule, or to an unwanted amount of a bioactive lipid which contributes to disease when present in excess.
- compositions and methods can be used to treat these ocular diseases and disorders, particularly by decreasing the effective in vivo concentration of a particular target lipid, for example, SlP and /or LPA.
- compositions and methods of the invention are useful in treating ocular diseases characterized, at least in part, by aberrant neovascularization, angiogenesis, fibrogenesis, fibrosis, scarring, inflammation, and immune response.
- Ischemic retinopathies are a diverse group of disorders characterized by a compromised retinal blood flow. Examples of IR include diabetic retinopathy (DR), retinopathy of prematurity (ROP), sickle cell retinopathy and retinal venous occlusive disease. AU of these disorders can be associated with a VEGF driven proliferation of pathological retinal neovascularization which can ultimately lead to intraocular hemorrhaging, epi-retinal membrane formation and tractional retinal detachment. Idiopathic epi-retinal membranes (ERMs), also called macular pucker or cellophane retinopathy, can cause a reduction in vision secondary to distortion of the retinal architecture.
- DR diabetic retinopathy
- ROP retinopathy of prematurity
- AU of these disorders can be associated with a VEGF driven proliferation of pathological retinal neovascularization which can ultimately lead to intraocular hemorrhaging, epi-retinal membrane formation and
- VEGF and its receptors are localized to ERMs.
- the presence of VEGF in membranes associated with proliferative diabetic retinopathy, proliferative vitreoretinopathy and macular pucker further suggests that this cytokine plays an important role in angiogenesis in ischemic retinal disease and in membrane growth in proliferative vitreoretinal disorders.
- VEGF receptors, VEGFRl and VEGFR2 are also identified on cells in ERMs. These data show that VEGF may be an autocrine and/or paracrine stimulator that may contribute to the progression of vascular and avascular ERMs.
- TGF- ⁇ Transforming growth factor- ⁇
- ERMs Transforming growth factor- ⁇
- PDR proliferative diabetic retinopathy
- bFGF basic fibroblastic growth factor
- an agent that binds, antagonizes, inhibits the effects or the production of SlP will be effective at suppressing pathologic retinal neovascularization in ischemic retinopathies and posterior segment diseases characterized by vascular or avascular ERM formation.
- ocular conditions characterized, at least in part, by aberrant neovascularization or angiogenesis include age-related macular degeneration, corneal graft rejection, neovascular glaucoma, contact lens overwear, infections of the cornea, including herpes simplex, herpes zoster and protozoan infection, pterygium, infectious uveitis, chronic retinal detachment, laser injury, sickle cell retinopathy, venous occlusive disease, choroidal neovascularization, retinal angiomatous proliferation, and idiopathic polypoidal choroidal vasculopathy.
- age-related macular degeneration including herpes simplex, herpes zoster and protozoan infection, pterygium, infectious uveitis, chronic retinal detachment, laser injury, sickle cell retinopathy, venous occlusive disease, choroidal neovascularization, retinal angiomatous proliferation, and id
- PVR Proliferative vitreoretinopathy
- PVR proliferative vitreoretinopathy
- PVR membranes include mainly retinal pigmented epithelial cells, fibroblasts, macrophages and vascular endothelial cells [Jerdan JA et al (1989), Ophthalmology, vol 96: 801-10 and Vidinova et al. (2005), Klin Monatsbl Augenheilkd; vol 222:568-571].
- cytokines including platelet derived growth factor (PDGF), transforming growth factor (TGF) beta, basic fibroblastic growth factor (bFGF), interleukin -6 (IL)-6 and interleukin-8 (EL)-8
- PDGF platelet derived growth factor
- TGF transforming growth factor
- bFGF basic fibroblastic growth factor
- IL interleukin -6
- EL interleukin-8
- Sphingosine -1-Phosphate is a bioactive lysolipid with pleotrophic effects. It is pro- angiogenic, pro inflammatory (stimulates the recruitment of macrophages and mast cells) and pro-fibrotic (stimulates scar formation). SlP generally stimulates cells to proliferate and migrate and is anti-apoptotic. SlP achieves these biologically diverse functions through its interactions with numerous cytokines and growth factors.
- SPHINGOMAB vascular endothelial growth factor
- VEGF vascular endothelial growth factor
- bFGF vascular endothelial growth factor
- IL-6 vascular endothelial growth factor
- IL-8 vascular endothelial growth factor-8
- Binding of SlP to the SlP 1 receptor can also increase PDGF production; therefore an agent that binds SlP would also be expected to diminish PDGF production [Milstien and Spiegel (2006), Cancer Cell, vol 9:148-150].
- in vitro SlP transforms human RPE cells into a myofibroblast-like phenotype similar to the type seen in PVR.
- Uveitis is an inflammatory disorder of the uveal tract of the eye. It can affect the front (anterior) or back (posterior) of the eye or both. It can be idiopathic or infectious in etiology and can be vision-threatening. Idiopathic uveitis has been associated with increased CD4+ expression in the anterior chamber. [Calder et al. (1999), Invest Ophthalmol Vis Sci, vol 40: 2019-24]. Data also suggests a pathologic role of the T lymphocyte and its chemoattractant IP-10 in the pathogenesis of uveitis [Abu El-Asrar (2004), Am J Ophthalmol, vol 138: 401-11].
- chemokines in acute anterior uveitis include macrophage inflammatory proteins, monocyte chemoattractant protein- 1 and JJL-8. These cytokines probably play a critical role in leukocyte recruitment in acute anterior uveitis. [Verma et al. (1997), Curr Eye Res; vol 16; 1202-8]. Given the profound and pleiotropic effects of the SlP signaling cascade, it is believed that SPHINGOMAB and other immune moieties that reduce the effective concentration of bioactive lipid would serve as an effective method of reducing or modulating the intraocular inflammation associated with uveitis.
- the corneal wound healing response is of particular relevance for refractive surgical procedures since it is a major determinant of safety and efficacy. These procedures are performed for the treatment of myopia, hyperopia and astigmatism.
- Laser in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK) are the most common refractive procedures however others have been developed in an attempt to overcome complications. These complications include overcorrection, undercorrection, regression and stromal opacification among others.
- a number of common complications are related to the healing response and have their roots in the biologic response to surgery.
- One of the greatest challenges in corneal biology is to promote tissue repair via regeneration rather than fibrosis. It is believed that the choice between regeneration and fibrosis lies in the control of fibroblast activation.
- myofibroblasts may appear in the subepithelial stroma 1-2 weeks after surgery or injury. Myofibroblasts are presumably derived from keratocytes under the influence of TGF- ⁇ [Jester et al (2003) Exp Eye Res, vol 77: 581-592]. Corneal haze and stromal scarring are characterized by reduced corneal transparency and may be associated with fibroblast and myofibroblast generation.
- TGF- ⁇ and PDGF are important in stimulating myofibroblast differentiation [Folger et al. (2001), Invest Ophthalmol Vis Sci; 42: 2534-2541]. Haze can be noted in the central interface after LASIK under certain circumstances. These include diffuse lamellar keratitis, donut-shaped flaps, and retention of epithelial debris at the interface. It is likely that each of these is associated with increased access of TGF- ⁇ from epithelial cells to the activated keratocytes. [Netto et al. (2005), Cornea, vol 24: 509-522].
- Regression is most likely due to heightened epithelial-stromal wound healing interactions such as increased production of epithelium modulating growth factors by corneal fibroblasts and or myofibroblasts [Netto et al. (2005), Cornea, vol 24: 509-522].
- Inhibition of TGF- ⁇ binding to receptors with topical anti-TGF- ⁇ antibody has been shown to reduce haze induced by PRK [Jester et al. (1997), Cornea, vol 16: 177-187].
- PRK topical anti-TGF- ⁇
- Glaucoma is classically thought of a disease whereby elevated intraocular pressure causes damage to the optic nerve and ultimately compromises the visual field and or the visual acuity.
- Other forms of glaucoma exist where optic nerve damage can occur in the setting of normal pressure or so called "normal tension glaucoma".
- glaucoma filtration surgery is needed whereby a fistula is surgically created in the eye to allow fluid to drain. This can be accomplished via trabeculectomy, the implantation of a medical device or other methods of surgical intervention. Glaucoma filtration surgery fails due to a wound healing process characterized by the proliferation of fibroblasts and ultimately scarring.
- SlP also stimulates the profibrotic function of multiple fibroblast cell types and the transformation into the myofibroblast phenotype and collagen production.
- an agent that binds, antagonizes, inhibits the effects or the production of SlP, or perhaps other bioactive lipids such as LPA will be effective at modulating the wound healing and/or fibrotic response that leads to failure of glaucoma surgery and will be an effective therapeutic method of enhancing successful surgical outcomes.
- the agent could be administered, e.g., via intravitreal or subconjunctival injection or topically.
- Corneal transplantation (penetrating keratoplasty (PK)) is the most successful tissue transplantation procedure in humans. Yet of the 47,000 corneal transplants performed annually in the United States, corneal allograft rejection is still the leading cause of corneal graft failure. [Ing JJ et al. (1998), Ophthalmology, vol 105: 1855-1865]. Currently, we do not sufficiently have the ability to avert allograft rejection although immunosuppression and immunomodulation may be a promising approach. Recently it has been discovered that CD4(+) T cells function as directly as effector cells and not helper cells in the rejection of corneal allografts. [Hegde S et al. (2005), Transplantation, vol 79: 23-31].
- Macrophages were the main infiltrating cell type followed by T-cells, mast cells and neutrophils.
- the early chemokine expression in high risk corneal transplantation was the mouse homologue of IL-8 (macrophage inflammatory protein-2) and monocyte chemotactic protein- 1 (MCP-I) [Yamagami S et al. (2005), MoI Vis, vol 11, 632-40].
- FTY720 is a novel immunosuppressive drug that acts by altering lymphocyte trafficking; resulting in peripheral blood lymphopenia and increased lymphocyte counts in lymph nodes. FTY mediates its immune-modulating effects by binding to some of the SP receptors expressed on lymphocytes. [Bohler T et al. (2005), Transplantation, vol 79: 492-5]. The drug is administered orally and a single oral dose reduced peripheral lymphocyte counts by 30-70%. FTY reduced T-cell subset, CD4(+) cells more than CD8(+) cells. [Bohler et al. (2004), Nephrol Dial Transplant, vol 19: 702-13].
- FTY treated mice showed a significant prolongation of orthotopic corneal-graft survival when administered orally. [Zhang et al. (2003), Transplantation, vol 76: 1511-3]. FTY oral treatment also significantly delayed rejection and decreased its severity in a rat-to-mouse model of corneal xenotransplantation [Sedlakova et al. (2005), Transplantation, vol 79, 297-303].
- immune moieties that decrease the effective concentration of bioactive lipids e.g., SPHINGOMAB
- SPHINGOMAB will also be useful in treatment of immunologic conditions such as allograft rejection, for example by attenuating the immune response, and thus will likely improve corneal graft survival after PK.
- the drug may also have the added advantage that in addition to systemic administration, local administration, e.g., via topical periocular or intraocular delivery, may be possible.
- Other ocular diseases with an inflammatory or immune component include chronic vitritis, infections, including herpes simplex, herpes zoster and protozoan infections, and ocular histoplasmosis.
- Treatment with an antibody targeted to bioactive lipid also is believed to benefit several conditions characterized by scarring of the anterior portion of the eye. These include the following:
- the cornea as the most anterior structure of the eye, is exposed to various hazards ranging from airborne debris to blunt trauma that can result in mechanical trauma.
- the cornea and anterior surface of the eye can also be exposed to other forms of trauma from surgery, and chemical, such as acid and alkali, injuries.
- the results of these types of injuries can be devastating often leading to corneal and conjunctival scarring symblephera formation.
- corneal neovascularization may ensue.
- Neutrophils accumulate, their release of leukotrienes, and the presence of interleukin-1 and interleukin-6, serves to recruit successive waves of inflammatory cells [Sotozono et al.
- OPC Ocular Cicatricial Pemphigoid
- OCP is a chronic cicatrizing (scar-forming) autoimmune disease that primarily affects the conjunctiva.
- the disease is invariably progressive and the prognosis is quite poor.
- conjunctival scarring and the associated keratopathy lead to bilateral blindness.
- Histologically the conjunctiva shows submucosal scarring and chronic inflammation in which mast cell participation is surprisingly great. [Yao L et al. (2003), Ocul Immunol Inflamm, vol 11: 211-222].
- Autoantigens lead to the formation of autoantibodies.
- SJS Stevens Johnson Syndrome
- TEN Toxic Epidermal Necrolysis
- SJS and TEN are life-threatening adverse reactions to medications.
- the ocular sequelae of these two related conditions can be severe and involve pathologic changes of the bulbar and palpebral conjunctiva, eyelids and cornea. Drugs and infections are the most common precipitating factors.
- Chronic eye findings include scarring, symblepharon formation, and cicatrisation of the conjunctiva as a result of the initial inflammatory process. This leads to entropion formation, trichiasis and instability of the tear film. Breakdown of the ocular surface leads to corneal scarring, neovascularization, and in severe cases keratinization. As in OCP subepithelial fibrosis of the conjunctiva occurs.
- SJS/TEN A vigorous autoimmune lymphocyte response to a drug or infection is believed to play a role in development of SJS/TEN.
- the infiltrating cell population in SJS includes macrophages, CD4 positive T cells, and CD8 positive T cells. This cell population is similar to those seen in chemical injury. [Kawasaki et al. (2000), J Ophthalmol, vol 84: 1191-3].
- a pterygium appears as a fleshy, vascular mass that occurs in the interpalpebral fissure.
- the body of the pterygium is a fleshy fibrovascular mass.
- Active pterygium are characterized by marked vascular engorgement and progressive growth. They are firmly adherent to the globe. In advanced cases the pterygium encroaches onto the cornea and may cause visual loss secondary to loss of corneal transparency within the visual axis or irregular astigmatism. Symptomatically, patients may experience foreign body sensation, tearing and blurred vision. Histopathology demonstrates hyalinization of the subepithelial connective tissue of the substantia limbal, increased number of fibroblasts and increased mast cells.
- ocular diseases and conditions with a fibrogenesis, fibrosis or scarring component include AMD, diabetic retinopathy, retinopathy of prematurity, sickle cell retinopathy, ischemic retinopathy, retinal venous occlusive disease and contact lens overwear.
- bioactive lipids like SlP and LPAs play a role in this process and an antibody-related treatment to diminish the concentrations of these agents will likely lead to therapeutic benefit to patients receiving the treatment.
- inhibitors of bioactive lipids particularly monoclonal antibodies directed against SlP and/or LPA, are believed to be useful in modulating surgical and traumatic wound healing responses.
- compositions and methods of the invention will be useful in treating disorders and diseases characterized, at least in part, by aberrant neovascularization, angiogenesis, fibrogenesis, fibrosis, scarring, inflammation, and immune response.
- scleroderma which is also referred to as systemic sclerosis.
- Scleroderma is an autoimmune disease that causes scarring or thickening of the skin, and sometimes involves other areas of the body, including the lungs, heart, and/or kidneys.
- Scleroderma is characterized by the formation of scar tissue (fibrosis) in the skin and organs of the body, which can lead to thickening and firmness of involved areas, with consequent reduction in function.
- SlP and LPA are pro-fibrotic growth factors that can contribute to fibroblast activation, proliferation, and the resulting increased fibroblast activity associated with maladaptive scarring and remodeling.
- S IP and LPA potential roles for S IP and LPA in activity of skin and other types of fibroblasts have been demonstrated.
- LPA stimulates the migration of murine skin fibroblasts (Hama, et al, J Biol Chem. 2004 Apr 23;279(17):17634-9), and human skin fibroblasts express several SlP receptor subtypes (Zhang, et al., Blood. 1999 May 1;93(9):2984- 90).
- SlP may have many potential indirect effects on fibroblast activity.
- SlP may facilitate the action of other well-known pro-fibrotic factors, such as TGF- ⁇ and platelet derived growth factor (PDGF).
- TGF- ⁇ is one of the most widely studied and recognized contributors to fibrosis (Desmouliere, et al., / Cell Biol 122: 103-111, 1993).
- TGF- ⁇ upregulates SphKl expression and activity leading to increased expression of tissue inhibitors of metalloproteinases 1 (TIMP-I), a protein that inhibits ECM degradation (Yamanaka, et a ⁇ ., JBiol Chem 279: 53994-54001, 2004).
- TIMP-I tissue inhibitors of metalloproteinases 1
- SlP stimulates expression and release of TGF- ⁇ (Norata, et al., Circulation 111: 2805-2811, 2005).
- S IP and PDGF There is also distinct evidence of crosstalk between S IP and PDGF.
- compositions and methods of the invention can be used to treat scleroderma, particularly by decreasing the effective in vivo concentration of a particular target lipid, for example, SlP and/or LPA.
- the treatment for diseases and conditions such as the examples given above can be administered by various routes employing different formulations and devices.
- Suitable pharmaceutically acceptable diluents, carriers, and excipients are well known in the art.
- One skilled in the art will appreciate that the amounts to be administered for any particular treatment protocol can readily be determined. Suitable amounts might be expected to fall within the range of 10 ⁇ g/dose to 10 g/dose, preferably within 10 mg/dose to 1 g/dose.
- Drug substances may be administered by techniques known in the art, including but not limited to systemic, subcutaneous, intradermal, mucosal, including by inhalation, and topical administration.
- the mucosa refers to the epithelial tissue that lines the internal cavities of the body.
- the mucosa comprises the alimentary canal, including the mouth, esophagus, stomach, intestines, and anus; the respiratory tract, including the nasal passages, trachea, bronchi, and lungs; and the genitalia.
- the mucosa will also include the external surface of the eye, i.e. the cornea and conjunctiva.
- Local administration (as opposed to systemic administration) may be advantageous because this approach can limit potential systemic side effects, but still allow therapeutic effect.
- compositions used in the present invention include, but are not limited to, solutions, emulsions, and liposome-containing formulations. These compositions may be generated from a variety of components that include, but are not limited to, preformed liquids, self -emulsifying solids and self-emulsifying semisolids.
- the pharmaceutical formulations used in the present invention may be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the active ingredients with the pharmaceutical carrier(s) or excipient(s).
- Preferred carriers include those that are pharmaceutically acceptable, particularly when the composition is intended for therapeutic use in humans.
- veterinarily acceptable carriers may be employed.
- the formulations are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.
- compositions of the present invention may be formulated into any of many possible dosage forms such as, but not limited to, tablets, capsules, liquid syrups, soft gels, suppositories, and enemas.
- the compositions of the present invention may also be formulated as suspensions in aqueous, non-aqueous or mixed media.
- Aqueous suspensions may further contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol and/or dextran.
- the suspension may also contain stabilizers.
- compositions may be formulated and used as foams.
- Pharmaceutical foams include formulations such as, but not limited to, emulsions, microemulsions, creams, jellies and liposomes.
- an immune-derived moiety can be delivered to the eye via, for example, topical drops or ointment, periocular injection, intracamerally into the anterior chamber or vitreous, via an implanted depot, or systemically by injection or oral administration.
- the quantity of antibody used can be readily determined by one skilled in the art.
- the traditional approaches to delivering therapeutics to the eye include topical application, redistribution into the eye following systemic administration or direct intraocular/periocular injections [Sultana et al. (2006),Current Drug Delivery, vol 3: 207-217; Ghate and Edelhauser (2006), Expert Opinion, vol 3: 275-287 and Kaur and Kanwar (2002), Drug Develop Industrial Pharmacy, vol 28: 473-493].
- Topical drops are convenient, but wash away primarily because of nasolacrimal drainage often delivering less than 5% of the applied drug into the anterior section of the eye and an even smaller fraction of that dose to the posterior segment of the globe.
- sprays afford another mode for topical administration.
- a third mode is ophthalmic ointments or emulsions can be used to prolong the contact time of the formulation with the ocular surface although blurring of vision and matting of the eyelids can be troublesome.
- Such topical approaches are still preferable, since systemic administration of therapeutics to treat ocular disorders exposes the whole body to the potential toxicity of the drug.
- Treatment of the posterior segment of the eye is medically important because age-related macular degeneration, diabetic retinopathy, posterior uveitis, and glaucoma are the leading causes of vision loss in the United States and other developed countries. [Myles et al. (2005), Adv Drug Deliv Rev; 57: 2063-79].
- the most efficient mode of drug delivery to the posterior segment is intravitreal injection through the pars plana.
- direct injections require a skilled medical practitioner to effect the delivery and can cause treatment-limiting anxiety in many patients.
- Periocular injections an approach that includes subconjunctival, retrobulbar, peribulbar and posterior subtenon injections, are somewhat less invasive than intravitreal injections. Repeated and long-term intravitreal injections may cause complications, such as vitreous hemorrhage, retinal detachment, or endophthalmitis.
- the anti-bioactive lipid antibody treatment might also be administered using one of the newer ocular delivery systems [Sultana et al. (2006), Current Drug Delivery, vol 3: 207-217 and Ghate and Edelhauser (2006), Expert Opinion, vol 3: 275-287], including sustained or controlled release systems, such as (a) ocular inserts (soluble, erodible, non-erodible or hydrogel-based), corneal shields, eg, collagen-based bandage and contact lenses that provide controlled delivery of drug to the eye, (b) in situ gelling systems that provide ease of administration as drops that get converted to gel form in the eye, thereby providing some sustained effect of drug in the eye, (c) vesicular systems such as liposomes, niosomes/discomes, etc., that offers advantages of targeted delivery, bio-compatibility and freedom from blurring of vision, (d) mucoadhesive systems that provide better retention in the eye, (e) prodrugs (T) penetration enhancers, (g
- transscleral iontophoresis (Eljarrat-Binstock and Domb (2006), Control Release, 110: 479-89] is an important advance and may offer an effective way to deliver antibodies to the posterior segment of the eye.
- excipients might also be added to the formulated antibody to improve performance of the therapy, make the therapy more convenient or to clearly ensure that the formulated antibody is used only for its intended, approved purpose.
- excipients include chemicals to control pH, antimicrobial agents, preservatives to prevent loss of antibody potency, dyes to identify the formulation for ocular use only, solubilizing agents to increase the concentration of antibody in the formulation, penetration enhancers and the use of agents to adjust isotonicity and/or viscosity.
- Inhibitors of, e.g., proteases could be added to prolong the half life of the antibody.
- the antibody is delivered to the eye by intravitreal injection in a solution comprising phosphate-buffered saline at a suitable pH for the eye.
- the antibody might also be chemically modified to yield a pro-drug that is administered in one of the formulations or devices previously described above.
- the active form of the antibody is then released by action of an endogenous enzyme.
- Possible ocular enzymes to be considered in this application are the various cytochrome p450s, aldehyde reductases, ketone reductases, esterases or N-acetyl- ⁇ -glucosamidases.
- Other chemical modifications to the antibody could increase its molecular weight, and as a result, increase the residence time of the antibody in the eye.
- pegylation Harris and Chess (2003), Nat Rev Drug Discov; 2: 214-21
- a process that can be general or specific for a functional group such as disulfide [Shaunak et al. (2006), Nat Chem Biol ; 2:312-3] or a thiol [Doherty et al. (2005), Bioconjug Chem; 16: 1291-8].
- mice Female C57BL6/J mice were subjected to laser-induced rupture of Bruch's membrane and administered either 0.5 ⁇ g of Sphingomab or an isotype-matched non-specific (NS) antibody diluted in 2 ⁇ of physiological saline. Mice were sacrificed 14 and 28 days after laser rupture.
- choroidal flatmounts of the sclera-choroid-RPE complex were prepared and stained for vasculature (R. communis agglutinin I; red) and pericytes (CD140b; green).
- Digital images were captured using an epifluorescence Zeiss Axioplan 2 with RGB Spot high-resolution digital camera and laser scanning confocal microscope (BioRad MRC 1024, BioRad Corporation, Temecula, CA).
- a z-series capture was used and the sum of lesion area throughout the z-series was multiplied by the z thickness (4 ⁇ m) to obtain the lesion volume.
- the sclera-choroid-RPE complex was stained with Masson's Trichrome.
- the sclera-choroid-RPE complex was embedded in paraffin and then serially sectioned at a thickness of 6 microns. Approximately 30 sections per lesion were evaluated. Quantitation of the volume of collagen deposition was calculated in the same manner as described for CNV lesion volume.
- FIG. 1 Captured digital images are evaluated morphometrically using ImageJ software (Research Services Branch, National Institutes of Health, Bethesda, MD).
- Figure IA shows that SPHINGOMAB dramatically attenuates choroidal neovascularization 14 and 28 days after laser- induced rupture of Bruch's membrane.
- Figure IB shows that SPHINGOMAB significantly reduces fibrosis associated with CNV lesion formation 28 days after laser-induced rupture of Bruch's membrane.
- Example 2 SPHINGOMAB inhibits neovascularization through multiple mechanisms including inhibition of endothelial cell migration and tube formation.
- SlP promotes the migration of human umbilical vein endothelial cells (HUVECs) and, in Matrigel and other assays, the formation of de novo BV formation in vitro [112]; SPHINGOMAB can neutralize these effects of SlP.
- HUVECs seeded onto GF-reduced Matrigel formed multiple capillary-like structures in the presence of SlP and failed to form capillary-like structures in the absence of SlP or when co-incubated with SPHINGOMAB and SlP.
- SPHINGOMAB inhibits neovascularization through multiple mechanisms including mitigation of the effects of SlP, VEGF and bFGF in vivo.
- mice were heparinized and injected with the fluorescent lectin, Isolectin B4-FITC, which binds to adhesion molecules expressed by vascular EC that form the growing BVs.
- the plugs were then excised, frozen in OCT, sectioned and viewed for FITC-stained BVs.
- Data in Figure 3 A suggest that SlP is a more potent stimulator of neovascularization in vivo than bFGF or VEGF[Lee, et al, (1999), Biochem Biophys Res Commun,. vol 264: 743-50], as evidenced by the vast amount of FITC-stained BVs in the plugs containing SlP compared to the plugs containing bFGF or VEGF.
- Endogenous SlP from the blood and surrounding tissue could supply a wound with pro- angiogenic stimuli.
- SPHINGOMAB Optimally stimulated plugs (Matrigel supplemented with 0.5 ⁇ g/mL bFGF or lOmg/mL VEGF) were implanted into mice. Mice received i.p. injections of 25mg/kg SPHINGOMAB or saline every 48 hrs starting 1 day prior to Matrigel implantation. Each treatment group (Matrigel, Matrigel plus GF or Matrigel plus GF and administered SPHINGOMAB) consisted of a minimum of 6 mice.
- mice were treated with heparin, injected with Isolectin B4-FITC, the plugs excised, embedded in OCT freezing medium and sectioned. Micro-vascular density was qualitatively accessed by lectin-FITC stained vessels as shown in Figure 3C. BV staining was sporadic in control (untreated) plugs, whereas the plugs containing bFGF or VEGF demonstrated significant evidence of vascularization. The plugs from mice treated with the SPHINGOMAB demonstrated a significant reduction in BV formation compared to the bFGF or VEGF plugs from saline-treated mice.
- SlP makes profound contributions to wound healing by activating fibroblast migration, proliferation and collagen production; SPHINGOMAB neutralizes these effects.
- SIP's ability to promote wound healing 1) SlP increased Swiss-3T3 fibroblast proliferation as measured by 3 H-thymidine incorporation using standard methods ( Figure 4A); 2) SlP promoted the migration of cardiac fibroblasts in a standard scratch wound healing assay.
- FIG. 4B SlP promoted collagen expression by cardiac fibroblasts isolated from transgenic mice possessing the collagen Ia GFP reporter, as indicated by immunofluorescence microscopy ( Figure 4C); and 4) SlP induced the differentiation of WI-38 lung fibroblasts into myofibroblasts, cells that are active in scar remodeling, as indicated by increased expression of myofibroblast marker protein, ⁇ -smooth muscle actin, using immunoblot analysis ( Figure 4D). In each of these assays, SPHINGOMAB neutralized S IP's. It is anticipated that ocular fibroblasts would respond similarly to SlP and SPHINGOMAB.
- Example 5 SlP promotes transformation of ocular epithelial cells and fibroblasts into contractile, scar tissue-producing myofibroblasts.
- Pathological tissue fibrosis is a primary, contributing factor in a number of ocular disorders, including: age-related macular degeneration, diabetic retinopathy, retinopathy of prematurity, proliferative vitreoretinopathy and consequences of glaucoma surgery.
- myofibroblasts fibrocontractile, scar tissue-producing cells that have been termed "myofibroblasts".
- myofibroblasts Normally, myofibroblasts are responsible for tissue repair as part of the wound healing response following injury.
- altered number and function of myofibroblasts are implicated in diseases characterized by pathological scar tissue formation in the liver, skin, lung, kidney, heart and eyes.
- RPE retinal pigmented epithelial
- myofibroblast transformation of ocular fibroblasts can result in abnormal scar tissue production after eye injury leading to subsequent vision loss.
- ⁇ -Smooth muscle actin ⁇ -SMA; a myofibroblast marker
- Collagen is one of the primary structural proteins that supports all tissues in the body and is one of the main components of scar tissue. In the non-pathological setting, total collagen content within tissue is maintained via a balance between collagen production by fibroblasts and degradation by certain enzymes. A number of disorders that involve increased levels of scar tissue result, in part, from physiological and molecular processes that inhibit degradation of collagen that is need for scar formation.
- Example 6 SPHINGOMAB inhibits inflammatory and immune cell infiltration
- Inflammation is the first response in the remodeling process [7]. It is triggered both by ischemia and by cellular damage and results in up-regulation of cytokine expression which stimulates the migration of macrophages and neutrophils to the injured area for phagocytosis of dead cells and to further up-regulate the inflammatory response [Jordan et al.(1999), Cardiovasc Res,, vol 43: 860-78]. Mast cells are also important cellular mediators of the inflammatory response. SlP released from mast cells is responsible for many of the adverse responses seen in experimental animal models of inflammation [Jolly et al (2004), J Exp Med,, vol 199: 959-70 and Jolly et al (2005), Blood,, vol 105: 4736-42].
- SPHINGOMAB dramatically attenuated the density of inflammatory macrophages (Figure 6A) and mast cells ( Figure 6B) suggesting that SPHINGOMAB may neutralize immune and inflammatory damages during AMD.
- Example 7 SPHINGOMAB is highly specific for SlP
- a competitive ELISA demonstrates SPHINGOMAB's specificity for SlP compared to other bioactive lipids.
- SPHINGOMAB demonstrated no cross-reactivity to sphingosine (SPH), the immediate metabolic precursor of SlP or lysophosphatidic acid (LPA), an important extracellular signaling molecule that is structurally and functionally similar to SlP.
- SPHINGOMAB did not recognize other structurally similar lipids and metabolites, including ceramide-1 -phosphate (ClP), dihydrosphingosine (DH-SPH), phosphatidyl serine (PS), phosphatidyl ethanolamine (PE), or sphingomyelin (SM).
- SPHINGOMAB did cross react with dihydrosphingosine- 1 -phosphate (DH-SlP) and, to a lesser extent, sphingosylphoryl choline (SPC) ( Figure 7).
- LPA LPA related diseases
- diseases which may involve excessive fibrosis LPA may have some direct fibrogenic effects by stimulating collagen gene expression and proliferation of fibroblasts [Chen, et al. (2006) FEBS Lett. 580(19):4737-45].
- an anti-LPA mAb may be useful in the treatment of fibrosis and diseases characterized by excessive fibroblast activity. These diseases include but are not limited to various ocular disorders, cardiac remodeling and heart failure and scleroderma.
- Antibodies directed against the bioactive lipid, LPA, that demonstrate good performance characteristics both in in vitro assays and in vivo would be highly useful in therapeutic and diagnostic applications.
- LPA Lysophosphatidic acid
- LPA Lysophosphatidic acid
- the presence of anti-LPA antibodies was determined by analyzing the sera of the immunized mice by ELISA at 3 time points after immunization.
- the immune response varied greatly between individual mice, both with respect to time of antibody response and levels attained.
- titer> 125,000 was observed in at least half of the mice. Five mice with the highest antibody titer were selected to initiate hybridoma cell line development.
- the mAbs from 6 individual cell lines were fully characterized for their specificity by competition ELISA using a series of analogues and for their potency in a panel of in vitro assays (Table 1).
- the majority of the mAbs exhibited specificity for LPA isoforms.
- the antibody affinity was estimated to be in the picomolar range. Further testing in animal models will determine whether these mAbs may provide the basis for promising therapies for LPA related diseases.
- the specificity of the anti-LPA mAbs was evaluated by determining the binding to a set of LPA variants and related biolipids such as distearoyl-phosphatidic acid, lysophosphatidylcholine, SlP, ceramide and ceramide-1-phosphate.
- the IC 50 and cross- reactivity of the 6 selected mAbs plus two additional mAbs (504B58-3F8 and 504B104) directed against different LPA-related compounds are summarized in Table 3.
- 18:1 LPA did not compete with mAb 63 bound to immobilized 18:1 LPA, yet it bound LPA with K d values in the picomolar range. Thus, while all six mAbs bound with similar affinities to LPA, five out of six mAbs exhibited effective and specific binding to 18:1 LPA.
- LPA or 18:0 LPA (0.1 ⁇ M) as coating antigen As coating antigen.
- MB maximal binding (expressed as OD450).
- LPA 18:0 LPA was captured on ELISA plates. Each competitor lipid (up to 10 ⁇ M) was serially diluted in BSA (1 mg/ml)-PBS and then incubated with the mAbs (3 nM). Mixtures were then transferred to LPA coated wells and the amount of bound antibody was measured with a secondary antibody. Data was normalized to maximum signal (A 450 ) and is expressed as percent inhibition. Assays were performed in triplicate for parameter analysis.
- DLPC l-Palmytoyl-Z-myristoyl-sn-glycero-S-phosphocholine.
- DASA 1,2-Distearoyl- sn -glycero-3 -phosphate ; 14:0 LPA: l-Myristoyl-2-hydroxy- sn -glycero-3 -phosphate 16:0 LPA: l-Plmytoyl-2-hydroxy- sn -glycero-3-phosphate 18:0 LPA: l-Stearoyl-2-hydroxy- sn -glycero-3-phosphate 18:1 LPA: l-Oleoyl-2-hydroxy- sn -glycero-3-phosphate 18:2 LPA: l-Linoleoyl-2-hydroxy- sn -glycero-3-phosphate 20:4 LPA: l-Arachidonoyl-2-hydroxy- sn -glycero-3-phosphate S
- Microtiter ELISA plates (Costar, Cat No. 3361) were coated with rabbit anti-mouse IgG, F(ab') 2 fragment specific antibody (Jackson, 315-005-047) diluted inlM Carbonate Buffer (pH 9.5) at 37 0 C for 1 h. Plates were washed with PBS and blocked with PBS/BSA/Tween-20 for 1 hr at 37 0 C. For the primary incubation, dilutions of non-specific mouse IgG or human IgG, whole molecule (used for calibration curve) and samples to be measured were added to the wells.
- Microtiter ELISA plates (Costar, Cat No. 3361) were coated with LPA-BSA diluted in IM Carbonate Buffer (pH 9.5) at 37 0 C for 1 h. Plates were washed with PBS (137 mM NaCl, 2.68 mM KCl, 10.1 mM Na 2 HPO 4 , 1.76 mM KH 2 PO 4 ; pH 7.4) and blocked with PBS/BSA/Tween-20 for 1 h at room temperature or overnight at 4 0 C.
- PBS 137 mM NaCl, 2.68 mM KCl, 10.1 mM Na 2 HPO 4 , 1.76 mM KH 2 PO 4 ; pH 7.4
- the samples to be tested were diluted at 0.4 ⁇ g/mL, 0.2 ⁇ g/mL, 0.1 ⁇ g/mL, 0.05 ⁇ g/mL, 0.0125 ⁇ g/mL, and 0 ⁇ g/mL and 100 ⁇ l added to each well. Plates were washed and incubated with 100 ⁇ l per well of HRP conjugated goat anti-mouse (1:20,000 dilution) (Jackson, cat No 115-035-003) for 1 h at room temperature. After washing, the enzymatic reaction was detected with tetramethylbenzidine (Sigma, cat No T0440) and stopped by adding 1 M H 2 SO 4 . The optical density (OD) was measured at 450nm using a Thermo Multiskan EX. Raw data were transferred to GraphPad software for analysis.
- mAbs The specificity of mAbs was tested in ELISA assays.
- Microtiter plates ELISA plates (Costar, Cat No. 3361) were coated with 18:0 LPA-BSA diluted in IM Carbonate Buffer (pH 9.5) at 37 0 C for 1 h. Plates were washed with PBS (137 mM NaCl, 2.68 mM KCl, 10.1 mM Na 2 HPO 4 , 1.76 mM KH 2 PO 4 ; pH 7.4) and blocked with PBS/BSA/Tween-20 at 37 0 C for 1 h or overnight at room temperature.
- PBS 137 mM NaCl, 2.68 mM KCl, 10.1 mM Na 2 HPO 4 , 1.76 mM KH 2 PO 4 ; pH 7.4
- anti-LPA mAb For the primary incubation 0.4 ⁇ g/mL anti-LPA mAb and designated amounts of (14:0, 16:0, 18:0, 18:1, 18:2 and 20:4) LPA, DSPA, 18:1 LPC (lysophosphatidylcholine), SlP, ceramide and ceramide-1 -phosphate were added to wells of the ELISA plates and incubated at 37 0 C for 1 h.
- Monoclonal antibodies were purified from culture supernatants by passing culture supernatants over protein A/G columns (Pierce, Cat.No 53133) at 0,.5 mL/min.
- Mobile phases consisted of IX Pierce IgG binding Buffer (Cat.No 21001 ) and 0.1 M glycine pH 2.7 (Pierce, Elution Buffer, Cat.No 21004).
- Antibody collections in 0.1 M glycine were diluted 10 % (v/v) with 1 M Phosphate Buffer, pH 8.0, to neutralize the pH.
- IgG 1 collections were pooled and dialyzed exhaustively against IX PBS (Pierce Slide-A-Lyzer Cassette, 3,500 MWCO, Cat.No 66382).
- Eluates were concentrated using Centricon YM-3(10,000 MWCO Amicon Cat.No 4203) by centrifugation for 1 h at 2,500 rcf.
- the antibody concentration was determined by quantitative ELISA as described above using a commercial myeloma IgG 1 stock solution as a standard.
- Heavy chain types of mAbs were determined by ELISA using Monoclonal Antibody Isotyping Kit (Sigma, ISO-2).
- LT 1009 includes three complementarity determining regions (each a "CDR") in each of the two light chain polypeptides and each of the two heavy chain polypeptides that comprise each antibody molecule.
- CDR complementarity determining regions
- the amino acid sequences for each of these six CDRs is provided immediately below ("VL” designates the variable region of the immunoglobulin light chain, whereas "VH” designates the variable region of the immunoglobulin heavy chain):
- LT1009 HC nucleotide sequence [SEQ ID NO: 7]:
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US12/091,890 US20090220523A1 (en) | 2005-10-28 | 2006-10-27 | Compositions and methods for the treatment and prevention of fibrotic, inflammatory and neovascularization conditions |
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CU20210101A7 (en) * | 2021-12-15 | 2023-07-12 | Ct Ingenieria Genetica Biotecnologia | POLYPEPTIDES THAT BIND PROANGIOGENIC GROWTH FACTORS |
Family Cites Families (291)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3773919A (en) | 1969-10-23 | 1973-11-20 | Du Pont | Polylactide-drug mixtures |
USRE30750E (en) | 1972-05-15 | 1981-09-29 | Cardiac Resuscitator Corporation | Cardiac resuscitator and monitoring apparatus |
US3953293A (en) | 1972-05-31 | 1976-04-27 | Takeda Chemical Industries, Ltd. | Process for the preparation of xylostasin |
US3984395A (en) | 1972-07-07 | 1976-10-05 | Schering Corporation | Method of isolating gentamicin C2a |
US3974137A (en) | 1973-02-07 | 1976-08-10 | Bristol-Myers Company | Process for the preparation of 1-[L-(-)-γ-amino-α-hydroxybutyryl]-kanamycin A (RD-1341A) |
US3959255A (en) | 1973-05-10 | 1976-05-25 | Roussel-Uclaf | Antibiotic aminoglycosides, and process of preparation |
NL7407483A (en) | 1973-06-05 | 1974-12-09 | ||
JPS5644076B2 (en) | 1973-07-12 | 1981-10-16 | ||
US4179337A (en) | 1973-07-20 | 1979-12-18 | Davis Frank F | Non-immunogenic polypeptides |
US3962429A (en) | 1973-08-01 | 1976-06-08 | Chugai Seiyaku Kabushiki Kaisha | Method for reducing side effects of aminoglycoside antibiotics and composition therefor |
US3953422A (en) | 1973-08-17 | 1976-04-27 | Smithkline Corporation | Deoxyglucose derivatives |
JPS554118B2 (en) | 1973-08-29 | 1980-01-29 | ||
US4170642A (en) | 1973-10-01 | 1979-10-09 | Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai | Derivatives of kanamycin A |
US3988316A (en) | 1973-12-19 | 1976-10-26 | Schering Corporation | Antibiotics sisomicin and verdamicin I and complex containing same |
US4011390A (en) | 1974-02-15 | 1977-03-08 | Schering-Plough Corporation | Semi-synthetic aminocyclitol aminoglycoside antibiotics and methods for the preparation thereof |
FR2263745B1 (en) | 1974-03-12 | 1977-12-02 | Roussel Uclaf | |
FR2263747B1 (en) | 1974-03-12 | 1977-12-02 | Roussel Uclaf | |
CA1046512A (en) | 1974-03-19 | 1979-01-16 | Peter J.L. Daniels | Derivatives of diaminocyclitols and method for their preparation |
CH630644A5 (en) | 1974-04-10 | 1982-06-30 | Takeda Chemical Industries Ltd | METHOD FOR PRODUCING DESOXYAMINOGLYCOSIDE ANTIBIOTICS. |
US4125707A (en) | 1974-05-28 | 1978-11-14 | Societa' Farmaceutici Italia S.P.A. | Protected pseudotrisaccharide intermediate for paromomycin and neomycin derivatives |
US4049498A (en) | 1974-06-05 | 1977-09-20 | Schering Corporation | Methods for the preparation of semi-synthetic aminocyclitol aminoglycoside antibiotics |
US4217446A (en) | 1974-10-26 | 1980-08-12 | Pfizer Inc. | ωAmino-2-hydroxyalkyl derivatives of aminoglycoside antibiotics |
US4051315A (en) | 1974-11-13 | 1977-09-27 | Bristol-Myers Company | 6"-Deoxykanamycin B and 6"-deoxytobramycin |
FR2290908A1 (en) | 1974-11-13 | 1976-06-11 | Roussel Uclaf | NEW AMINOGLYCOSIDIC DERIVATIVE AND ITS SALTS, THEIR METHOD OF PREPARATION AND THEIR APPLICATION AS MEDICINAL PRODUCTS |
US4012576A (en) | 1974-12-12 | 1977-03-15 | Bristol-Myers Company | Antibiotic complex Bu 2183 |
US3978214A (en) | 1975-05-02 | 1976-08-31 | Schering Corporation | Novel 4,6-di-o-(aminoglycosyl)-2-deoxystreptamine, method for its manufacture, method for its use as an antiprotozoal agent and compositions useful thereof |
US3997524A (en) | 1975-05-02 | 1976-12-14 | Schering Corporation | Process for the manufacture of 6'-N-alkyl derivatives of sisomicin and verdamicin; novel intermediates useful therein, and novel 6'-N-alkylverdamicins prepared thereby |
US4009328A (en) | 1975-05-02 | 1977-02-22 | Schering Corporation | Aminoglycoside 66-40C, method for its manufacture, method for its use as an intermediate in the preparation of known antibiotics and novel antibacterials |
US4044123A (en) | 1975-05-02 | 1977-08-23 | Schering Corporation | 6'-N-alkyl-4,6-di-O-(aminoglycosyl)-1,3-diaminocyclitols, methods for their use as antibacterial agents and compositions useful therefor |
US3984393A (en) | 1975-09-08 | 1976-10-05 | The Upjohn Company | Aminoglycoside antibiotics |
US3996205A (en) | 1975-09-08 | 1976-12-07 | The Upjohn Company | Aminoglycoside antibiotics and intermediates |
US4038478A (en) | 1975-09-08 | 1977-07-26 | The Upjohn Company | O-glycoside ortho esters of neamine containing compounds |
US4002608A (en) | 1975-11-04 | 1977-01-11 | Schering Corporation | 1-N-alkyl-aminoglycoside-XK-88 derivatives and methods for their manufacture |
US4003922A (en) | 1975-11-24 | 1977-01-18 | Bristol-Myers Company | Synthesis of cis-1,4-cyclohexadiene dioxide |
JPS5265250A (en) | 1975-11-24 | 1977-05-30 | Bristol Myers Co | Whole synthesizing process for antimicrobial agents |
US4195170A (en) | 1975-12-09 | 1980-03-25 | Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai | 3',4'-Episulfido kanamycin B compounds |
GB1529243A (en) | 1976-02-25 | 1978-10-18 | Pfizer Ltd | Process for the preparation of aminoglycoside derivatives and cyclic urethane intermediates for use therein |
GB1573212A (en) * | 1976-04-15 | 1980-08-20 | Technicon Instr | Immunoassay for gentamicin |
PT66654B (en) | 1976-06-10 | 1978-11-10 | Canas Rodriguez Antonio | Process for the preparation of synthetic aminoglicosides |
JPS52153942A (en) | 1976-06-16 | 1977-12-21 | Microbial Chem Res Found | Preparation of kanamicin c deoxy derivatives and kanamicine c or its deoxy derivatives |
NZ184213A (en) | 1976-06-16 | 1980-05-27 | Pfizer | 2-deoxystreptamine aminoglycosides and antibacterial pharmaceutical compositions |
US4283528A (en) | 1976-06-17 | 1981-08-11 | Schering Corporation | 1-N-aminohydroxyacyl derivatives of gentamicin B |
US4136254A (en) | 1976-06-17 | 1979-01-23 | Schering Corporation | Process of selectively blocking amino functions in aminoglycosides using transition metal salts and intermediates used thereby |
US4230847A (en) | 1976-06-17 | 1980-10-28 | Schering Corporation | Aminoglycoside antibiotic compounds |
US4085208A (en) | 1976-06-21 | 1978-04-18 | Schering Corporation | Process for preparing 4,6-di-O-(aminoglycosyl)-1,3-diaminocyclitols and novel 1-epimers and 1-N-alkyl derivatives produced thereby; methods for the use of the 1-epimer derivatives as antibacterial agents and compositions useful therefor |
US4066752A (en) | 1976-06-21 | 1978-01-03 | Schering Corporation | 1-Desamino-1-hydroxy and 1-desamino-1-epi-hydroxy-4,6-di-o-(aminoglycosyl)-1,3-diaminocyclitols; 1-desamino-1-oxo-4,6-di-o-(aminoglycosyl)-1,3-diaminocyclitols, intermediates and use as antibacterial agents |
US4032404A (en) | 1976-07-14 | 1977-06-28 | Bristol-Myers Company | Fermentation process for producing apramycin and nebramycin factor V' |
US4146617A (en) | 1976-07-15 | 1979-03-27 | Roussel Uclaf | Desoxystreptamine derivatives, salts, pharmaceutical compositions and method of use |
US4189569A (en) | 1976-09-08 | 1980-02-19 | Abbott Laboratories | Seldomycin factor 5 derivatives |
US4187372A (en) | 1976-09-08 | 1980-02-05 | Abbott Laboratories | Seldomycin factor 5 derivative |
JPS5356661A (en) | 1976-10-28 | 1978-05-23 | Shionogi & Co Ltd | Novel amino glycoside antibiotics derivatives |
US4170643A (en) | 1977-04-13 | 1979-10-09 | Labaz | Aminoglycoside-aminocyclitol derivatives and method of use |
US4347354A (en) | 1977-04-28 | 1982-08-31 | Bristol-Myers Company | Preparation of 1-N-[ω-amino-α-hydroxyalkanoyl]aminoglycoside polysilylated antibiotics and products obtained therefrom |
US4424343A (en) | 1977-04-28 | 1984-01-03 | Bristol Myers Company | Preparation of 1-N- ω-amino-α-hydroxyalkanoyl!kanamycin polysilylates and products |
US4140849A (en) | 1977-05-23 | 1979-02-20 | Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai | Kanamycin C derivatives |
US4212859A (en) | 1977-06-24 | 1980-07-15 | Schering Corporation | 2'-Hydroxy-2'-desamino-4,6-di-O-(aminoglycosyl)-1,3-diaminocyclitols, methods for their manufacture, method for their use as antibacterial agents, and compositions useful therefor |
DK308878A (en) | 1977-08-18 | 1979-02-19 | Pfizer | AMINOGLYOSIDE DERIVATIVES AND PROCEDURES FOR THEIR PREPARATION |
CA1105455A (en) | 1977-09-19 | 1981-07-21 | Junji Irisawa | Aminoglycoside derivatives |
JPS5461151A (en) | 1977-10-25 | 1979-05-17 | Shionogi & Co Ltd | Novel aminoglycoside derivative |
US4214076A (en) | 1977-12-21 | 1980-07-22 | Abbott Laboratories | 2'-N-Substituted fortimicin B and derivatives |
US4187297A (en) | 1977-12-21 | 1980-02-05 | Abbott Laboratories | 3-De-O-methyl-2-N-acyl and alkyl fortimicins A and B |
US4196197A (en) | 1977-12-21 | 1980-04-01 | Abbott Laboratories | 2'N-Acyl and alkyl-6'-N-alkyl- and 6',6'-di-N-alkyl derivatives of fortimicins A and B |
US4176178A (en) | 1977-12-21 | 1979-11-27 | Abbott Laboratories | 2-Deoxy-2'-N-acyl and alkyl fortimicins A and B |
US4205070A (en) | 1977-12-21 | 1980-05-27 | Abbott Laboratories | 6'N-Alkyl- and 6',6'-di-N-alkyl derivatives of fortimicins A and B |
US4319022A (en) | 1977-12-21 | 1982-03-09 | Abbott Laboratories | 2-O-Substituted sulfonyl derivatives of fortimicin B |
US4192867A (en) | 1977-12-21 | 1980-03-11 | Abbott Laboratories | 2-Deoxyfortimicin A, 4-N-alkyl and 4-N-acyl-2-deoxyfortimicin B derivatives |
US4187296A (en) | 1977-12-21 | 1980-02-05 | Abbott Laboratories | 2-N-acyl and alkyl 6-epi-fortimicin B and derivatives |
US4214075A (en) | 1977-12-21 | 1980-07-22 | Abbott Laboratories | 6'-Epi-fortimicin A and B derivatives |
US4207314A (en) | 1977-12-21 | 1980-06-10 | Abbott Laboratories | Isofortimicin |
US4187298A (en) | 1977-12-21 | 1980-02-05 | Abbott Laboratories | 2'N-acyl and alkyl fortimicin B and derivatives, 4,2'-N,N'diacyl and dialkyl fortimicin B derivatives 4-N-acyl-2'-N-alkyl and 4-N-alkyl-2'-N-acyl fortimicin B derivatives |
US4169198A (en) | 1977-12-21 | 1979-09-25 | Abbott Laboratories | 2-Deoxyfortimicin B |
US4317904A (en) | 1977-12-21 | 1982-03-02 | Abbott Laboratories | 1,2-Epiminofortimicin B |
US4183920A (en) | 1977-12-21 | 1980-01-15 | Abbott Laboratories | 4-N-Acyl, 2'-N-acyl and 4,2'-N,N'-diacylfortimicin E derivatives |
US4187299A (en) | 1977-12-21 | 1980-02-05 | Abbott Laboratories | Fortimicin E |
JPS5492951A (en) | 1977-12-29 | 1979-07-23 | Shionogi & Co Ltd | Novel aminoglycoside derivative |
USRE30985E (en) | 1978-01-01 | 1982-06-29 | Serum-free cell culture media | |
JPS6030320B2 (en) | 1978-01-13 | 1985-07-16 | 山之内製薬株式会社 | New antibiotics and their production methods |
US4223022A (en) | 1978-01-16 | 1980-09-16 | Schering Corporation | Stabilized aminoglycoside antibiotic formulations |
US4226978A (en) | 1978-03-13 | 1980-10-07 | Miles Laboratories, Inc. | β-Galactosyl-umbelliferone-labeled aminoglycoside antibiotics and intermediates in their preparation |
US4275149A (en) | 1978-11-24 | 1981-06-23 | Syva Company | Macromolecular environment control in specific receptor assays |
US4318980A (en) | 1978-04-10 | 1982-03-09 | Miles Laboratories, Inc. | Heterogenous specific binding assay employing a cycling reactant as label |
US4181797A (en) | 1978-07-10 | 1980-01-01 | Bristol-Myers Company | 1-N-(ω-amino-α-hydroxyalkanoyl) derivatives of 4'-deoxy-6'-N-methylkanamycin A |
FR2435481A1 (en) | 1978-09-06 | 1980-04-04 | Roussel Uclaf | NOVEL AMINOGLYCOSIDES DERIVED FROM DESOXYSTREPTAMINE AND SALTS THEREOF, PROCESS FOR THE PREPARATION THEREOF AND APPLICATION AS MEDICAMENTS |
JPS5538345A (en) | 1978-09-11 | 1980-03-17 | Shionogi & Co Ltd | Novel aminoglycoside derivative |
JPS5540621A (en) | 1978-09-14 | 1980-03-22 | Shionogi & Co Ltd | Novel aminoglycoside derivative |
SE447386B (en) | 1978-10-18 | 1986-11-10 | Kowa Co | AMINOGLYCOSIDES, COMPOSITION CONTAINING THEM, AND PROCEDURE FOR PREPARING THEM |
IE48972B1 (en) | 1978-11-11 | 1985-06-26 | Microbial Chem Res Found | The production of a selectively protected n-acylated derivative of an aminoglycosidic antibiotic |
HU179145B (en) | 1979-02-01 | 1982-08-28 | Chinoin Gyogyszer Es Vegyeszet | Process for microbiological producing sysomycin with fermenting micromonospora danubiensis |
US4207415A (en) | 1979-02-05 | 1980-06-10 | Abbott Laboratories | Method of producing 2-deoxyfortimicin A |
US4208407A (en) | 1979-02-05 | 1980-06-17 | Abbott Laboratories | 5-Deoxyfortimicin A, 2,5-dideoxyfortimicin A and the corresponding 4-N-acyl and alkyl fortimicin B derivatives thereof and intermediates therefor |
HU179146B (en) | 1979-02-06 | 1982-08-28 | Chinoin Gyogyszer Es Vegyeszet | Process for microbiological producing sysnycin with fermenting micromonospora rosea |
US4223024A (en) | 1979-03-12 | 1980-09-16 | Abbott Laboratories | 4"-O-Alkylgentamicins and sagamicins |
US4213974A (en) | 1979-03-29 | 1980-07-22 | Abbott Laboratories | 4-N,2'-N and 4,2'-Di-N-fortimicin AO derivatives |
US4219643A (en) | 1979-03-29 | 1980-08-26 | Abbott Laboratories | Fortimicin AN |
US4219644A (en) | 1979-03-29 | 1980-08-26 | Abbott Laboratories | Fortimicins AH and AI |
US4214079A (en) | 1979-03-29 | 1980-07-22 | Abbott Laboratories | 4-N, 2'-N and 4,2'-Di-N-fortimicin AL derivatives |
US4330673A (en) | 1979-03-29 | 1982-05-18 | Abbott Laboratories | Process for producing 3-O-demethylaminoglycoside and novel 3-O-demethylfortimicin derivatives |
US4214080A (en) | 1979-03-29 | 1980-07-22 | Abbott Laboratories | Fortimicins AM and AP |
US4220756A (en) | 1979-03-29 | 1980-09-02 | Abbott Laboratories | Method of producing 3-O-demethylfortimicin B,4-N-alkylfortimicin B derivatives and related aminoglycoside antibiotics |
US4213972A (en) | 1979-03-29 | 1980-07-22 | Abbott Laboratories | 4-N, 2'-N and 4,2'Di-N-fortimicins AH and AI |
US4219642A (en) | 1979-03-29 | 1980-08-26 | Abbott Laboratories | Fortimicin AO |
US4216210A (en) | 1979-03-29 | 1980-08-05 | Abbott Laboratories | Fortimicins AM and AP derivatives |
US4213971A (en) | 1979-03-29 | 1980-07-22 | Abbott Laboratories | 4-N, 2'-N and 4,2'-Di-N-fortimicin AD derivatives |
US4214078A (en) | 1979-03-29 | 1980-07-22 | Abbott Laboratories | Fortimicin AL |
FR2452932A1 (en) | 1979-04-04 | 1980-10-31 | Toyo Jozo Kk | NOVEL AMINOGLYCOSIDE ANTIBIOTICS AND THEIR PRODUCTION |
JPS5629598A (en) | 1979-08-18 | 1981-03-24 | Toyo Jozo Co Ltd | Novel amino sugar antibiotic g-367-2 and its preparation |
DE2921973A1 (en) | 1979-05-30 | 1980-12-11 | Bayer Ag | SISOMICIN DERIVATIVES, METHOD FOR THE PRODUCTION THEREOF AND THEIR USE AS MEDICINAL PRODUCTS |
DE2921974A1 (en) | 1979-05-30 | 1980-12-11 | Bayer Ag | SELECTIVELY PROTECTED 1-N- (OMEGA-AMINOALKYLOXYCARBONYL) -SISOMICIN DERIVATIVES |
US4250170A (en) | 1979-06-11 | 1981-02-10 | Bristol-Myers Company | Antibacterial agents Bu-2349A and B and method of using same |
JPS6023084B2 (en) | 1979-07-11 | 1985-06-05 | 味の素株式会社 | blood substitute |
US4252972A (en) | 1979-09-26 | 1981-02-24 | Abbott Laboratories | Fortimicin B-1,2:4,5-bis-carbamates |
US4251516A (en) | 1979-09-26 | 1981-02-17 | Abbott Laboratories | 2-Deoxy-3-O-Demethylfortimicins |
US4250304A (en) | 1979-09-26 | 1981-02-10 | Abbott Laboratories | 2-Deoxy-2-substituted fortimicin A and B and derivatives |
US4251511A (en) | 1979-10-02 | 1981-02-17 | The Upjohn Company | Antibiotic and fermentation process of preparing |
US4288547A (en) | 1979-11-13 | 1981-09-08 | Haruo Yamamoto | Fermentative process for preparing antibiotics of the gentamicin class |
US4387219A (en) | 1979-11-13 | 1983-06-07 | Sterling Drug Inc. | 2-Hydroxy gentamicin compounds |
DE3000841A1 (en) | 1980-01-11 | 1981-07-16 | Bayer Ag, 5090 Leverkusen | INSULATION AND CLEANING OF AMINOGLYCOSIDE ANTIBIOTIAKA |
JPS56118097A (en) | 1980-02-25 | 1981-09-16 | Microbial Chem Res Found | Kanamycin a derivative and its preparation |
US4376110A (en) | 1980-08-04 | 1983-03-08 | Hybritech, Incorporated | Immunometric assays using monoclonal antibodies |
US4468513A (en) | 1980-09-22 | 1984-08-28 | Eli Lilly And Company | 2'-N-Acylated and 2'-N-alkylated derivatives of 4-O-substituted-2-deoxystreptamine aminoglycosides |
US4424344A (en) | 1980-09-22 | 1984-01-03 | Eli Lilly And Company | 2-N-Acylated and 2-N-alkylated derivatives of 4-O-substituted-2-deoxystreptamine aminoglycosides and process |
US4468512A (en) | 1980-09-22 | 1984-08-28 | Eli Lilly And Company | 1-N-Acylated and 1-N-alkylated derivatives of 4-O-substituted-2-deoxystreptamine aminoglycosides |
US4424345A (en) | 1980-09-22 | 1984-01-03 | Eli Lilly And Company | 1-N-Acylated and 1-N-alkylated derivatives of 4-O-substituted-2-deoxystreptamine aminoglycosides and process |
DE3101376A1 (en) | 1981-01-17 | 1982-09-02 | Bayer Ag, 5090 Leverkusen | SISOMICIN DERIVATIVES, METHOD FOR THE PRODUCTION THEREOF AND THEIR USE AS MEDICINAL PRODUCTS |
DE3206725A1 (en) | 1981-05-13 | 1982-12-02 | Merck Patent Gmbh, 6100 Darmstadt | PERSONALLY SOLUBLE SALTS OF AMINOGLYCOSIDANTIBIOTICS |
US4485045A (en) | 1981-07-06 | 1984-11-27 | Research Corporation | Synthetic phosphatidyl cholines useful in forming liposomes |
JPS5815994A (en) | 1981-07-22 | 1983-01-29 | Microbial Chem Res Found | Kanamycin derivative modified at 2' position and its preparation |
JPS5821692A (en) | 1981-07-29 | 1983-02-08 | Microbial Chem Res Found | Novel aminoglycoside |
FI822671L (en) | 1981-08-07 | 1983-02-08 | Sandoz Ag | AMINOGLYCOSIDE DERIVATIVES, PHARMACEUTICAL COMPOSITION FOR FRAMSTAELLNING AV DEM, PHARMACEUTICAL COMPOSITION INNEHAOLLANDE DEM |
US4640835A (en) | 1981-10-30 | 1987-02-03 | Nippon Chemiphar Company, Ltd. | Plasminogen activator derivatives |
US4418193A (en) | 1982-04-09 | 1983-11-29 | Abbott Laboratories | Method of producing 2-epi-fortimicin A |
JPS58213774A (en) | 1982-06-04 | 1983-12-12 | Kowa Co | Novel aminoglycoside |
US4645760A (en) | 1982-07-30 | 1987-02-24 | Health Research Inc. | Activated aminoglycosides and aminoglycoside-aminocyclitols pharmaceutical compositions and method of use |
US4493831A (en) | 1982-10-25 | 1985-01-15 | Fujisawa Pharmaceutical Co., Ltd. | Aminoglycoside derivatives |
US4560655A (en) | 1982-12-16 | 1985-12-24 | Immunex Corporation | Serum-free cell culture medium and process for making same |
US4657866A (en) | 1982-12-21 | 1987-04-14 | Sudhir Kumar | Serum-free, synthetic, completely chemically defined tissue culture media |
AU575038B2 (en) | 1983-01-28 | 1988-07-21 | Fujisawa Pharmaceutical Co., Ltd. | Aminoglycoside derivatives, eg 2'n-palmitoyl-kasugamycin |
US4568649A (en) | 1983-02-22 | 1986-02-04 | Immunex Corporation | Immediate ligand detection assay |
GB8308235D0 (en) | 1983-03-25 | 1983-05-05 | Celltech Ltd | Polypeptides |
US4816567A (en) | 1983-04-08 | 1989-03-28 | Genentech, Inc. | Recombinant immunoglobin preparations |
US4544545A (en) | 1983-06-20 | 1985-10-01 | Trustees University Of Massachusetts | Liposomes containing modified cholesterol for organ targeting |
JPS6042394A (en) | 1983-08-18 | 1985-03-06 | Kowa Co | Novel aminoglycoside and its preparation |
US4767704A (en) | 1983-10-07 | 1988-08-30 | Columbia University In The City Of New York | Protein-free culture medium |
US4626513A (en) | 1983-11-10 | 1986-12-02 | Massachusetts General Hospital | Method and apparatus for ligand detection |
US4496689A (en) | 1983-12-27 | 1985-01-29 | Miles Laboratories, Inc. | Covalently attached complex of alpha-1-proteinase inhibitor with a water soluble polymer |
ATE56096T1 (en) | 1984-03-15 | 1990-09-15 | Immunex Corp | TEST FOR INSTANT DETECTION OF LIGANDS, TEST KIT AND ITS PREPARATION. |
US4658830A (en) | 1984-08-08 | 1987-04-21 | Survival Technology, Inc. | Method and apparatus for initiating reperfusion treatment by an unattended individual undergoing heart attack symptoms |
GB8422238D0 (en) | 1984-09-03 | 1984-10-10 | Neuberger M S | Chimeric proteins |
US4879231A (en) | 1984-10-30 | 1989-11-07 | Phillips Petroleum Company | Transformation of yeasts of the genus pichia |
US4656160A (en) | 1984-11-29 | 1987-04-07 | Fujisawa Pharmaceutical Co., Ltd. | Aminoglycoside derivatives |
US5506337A (en) | 1985-03-15 | 1996-04-09 | Antivirals Inc. | Morpholino-subunit combinatorial library and method |
US4737456A (en) | 1985-05-09 | 1988-04-12 | Syntex (U.S.A.) Inc. | Reducing interference in ligand-receptor binding assays |
US4895724A (en) | 1985-06-07 | 1990-01-23 | Pfizer Inc. | Chitosan compositions for controlled and prolonged release of macromolecules |
EP0206448B1 (en) | 1985-06-19 | 1990-11-14 | Ajinomoto Co., Inc. | Hemoglobin combined with a poly(alkylene oxide) |
GB8516415D0 (en) | 1985-06-28 | 1985-07-31 | Celltech Ltd | Culture of animal cells |
JPS6253997A (en) | 1985-09-03 | 1987-03-09 | Kowa Co | Novel amino glycoside and pharmaceutical preparation containing same |
US4676980A (en) | 1985-09-23 | 1987-06-30 | The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services | Target specific cross-linked heteroantibodies |
IT1200774B (en) | 1985-10-10 | 1989-01-27 | Pierrel Spa | AMIKACINA FEELING PROCEDURE |
US5008288A (en) | 1986-01-06 | 1991-04-16 | Alfred Stracher | Carnitine directed pharmaceutical agents |
US5225539A (en) | 1986-03-27 | 1993-07-06 | Medical Research Council | Recombinant altered antibodies and methods of making altered antibodies |
GB8607679D0 (en) | 1986-03-27 | 1986-04-30 | Winter G P | Recombinant dna product |
US6548640B1 (en) | 1986-03-27 | 2003-04-15 | Btg International Limited | Altered antibodies |
US4927762A (en) | 1986-04-01 | 1990-05-22 | Cell Enterprises, Inc. | Cell culture medium with antioxidant |
GB8610600D0 (en) | 1986-04-30 | 1986-06-04 | Novo Industri As | Transformation of trichoderma |
US4791192A (en) | 1986-06-26 | 1988-12-13 | Takeda Chemical Industries, Ltd. | Chemically modified protein with polyethyleneglycol |
US5260203A (en) | 1986-09-02 | 1993-11-09 | Enzon, Inc. | Single polypeptide chain binding molecules |
US4946778A (en) | 1987-09-21 | 1990-08-07 | Genex Corporation | Single polypeptide chain binding molecules |
US5869620A (en) | 1986-09-02 | 1999-02-09 | Enzon, Inc. | Multivalent antigen-binding proteins |
US5567610A (en) | 1986-09-04 | 1996-10-22 | Bioinvent International Ab | Method of producing human monoclonal antibodies and kit therefor |
US4816450A (en) * | 1986-09-15 | 1989-03-28 | Duke University | Inhibition of protein kinase C by long-chain bases |
US4937232A (en) * | 1986-09-15 | 1990-06-26 | Duke University | Inhibition of protein kinase C by long-chain bases |
JPH0764866B2 (en) | 1987-02-24 | 1995-07-12 | 財団法人微生物化学研究会 | 1-N- (4-amino-3-fluoro-2-hydroxybutyryl) kanamycin |
WO1988007089A1 (en) | 1987-03-18 | 1988-09-22 | Medical Research Council | Altered antibodies |
US5204244A (en) | 1987-10-27 | 1993-04-20 | Oncogen | Production of chimeric antibodies by homologous recombination |
IT1225484B (en) | 1987-11-27 | 1990-11-14 | Pierrel Spa | SYNTHESIS PROCEDURE OF AMIKACINA |
DE3741056A1 (en) * | 1987-12-04 | 1989-08-24 | Behringwerke Ag | MANUMYCINE DERIVATIVES, METHOD FOR THEIR PRODUCTION AND THEIR USE |
US5010175A (en) | 1988-05-02 | 1991-04-23 | The Regents Of The University Of California | General method for producing and selecting peptides with specific properties |
US5030723A (en) | 1988-05-31 | 1991-07-09 | The Biomembrane Institute | Long-chain glycolipid structure |
ATE135397T1 (en) | 1988-09-23 | 1996-03-15 | Cetus Oncology Corp | CELL CULTIVATION MEDIUM FOR INCREASED CELL GROWTH, TO INCREASE THE LONGEVITY AND EXPRESSION OF THE PRODUCTS |
GB8823869D0 (en) | 1988-10-12 | 1988-11-16 | Medical Res Council | Production of antibodies |
US5175384A (en) | 1988-12-05 | 1992-12-29 | Genpharm International | Transgenic mice depleted in mature t-cells and methods for making transgenic mice |
US20040049014A1 (en) | 1988-12-28 | 2004-03-11 | Protein Design Labs, Inc. | Humanized immunoglobulins |
US5530101A (en) | 1988-12-28 | 1996-06-25 | Protein Design Labs, Inc. | Humanized immunoglobulins |
US5442047A (en) | 1991-12-04 | 1995-08-15 | Schering Corporation | Process for preparing isepamicin |
DE3920358A1 (en) | 1989-06-22 | 1991-01-17 | Behringwerke Ag | BISPECIFIC AND OLIGO-SPECIFIC, MONO- AND OLIGOVALENT ANTI-BODY CONSTRUCTS, THEIR PRODUCTION AND USE |
IT1237490B (en) | 1989-09-22 | 1993-06-07 | Chementecno S R L Monza Milano | PROCEDURE FOR THE SYNTHESIS OF 1 N (S DELTA AMINO ALFA IDROSSIBUTIRRIL) KAMANYCIN A BASED ON THE TREATMENT OF 1 N (S DELTA BENZYOXYCARBONLAMINE ALPHA HYDROXYBUTYRRIL) 3.6 'OF N BENZYLSYCARBONYLKANAMYCIN A WITH FORMIC ACID |
US5013556A (en) | 1989-10-20 | 1991-05-07 | Liposome Technology, Inc. | Liposomes with enhanced circulation time |
US6673986B1 (en) | 1990-01-12 | 2004-01-06 | Abgenix, Inc. | Generation of xenogeneic antibodies |
US6150584A (en) | 1990-01-12 | 2000-11-21 | Abgenix, Inc. | Human antibodies derived from immunized xenomice |
US6075181A (en) | 1990-01-12 | 2000-06-13 | Abgenix, Inc. | Human antibodies derived from immunized xenomice |
WO1991010741A1 (en) | 1990-01-12 | 1991-07-25 | Cell Genesys, Inc. | Generation of xenogeneic antibodies |
ATE111917T1 (en) | 1990-03-08 | 1994-10-15 | Opos Biochimica Srl | METHODS OF PRODUCTION OF AMIKACIN PRECURSORS. |
US5229275A (en) | 1990-04-26 | 1993-07-20 | Akzo N.V. | In-vitro method for producing antigen-specific human monoclonal antibodies |
US5270163A (en) | 1990-06-11 | 1993-12-14 | University Research Corporation | Methods for identifying nucleic acid ligands |
ES2259800T3 (en) | 1990-06-11 | 2006-10-16 | Gilead Sciences, Inc. | PROCEDURES FOR THE USE OF NUCLEIC ACID LINKS. |
IE66205B1 (en) | 1990-06-14 | 1995-12-13 | Paul A Bartlett | Polypeptide analogs |
US5650489A (en) | 1990-07-02 | 1997-07-22 | The Arizona Board Of Regents | Random bio-oligomer library, a method of synthesis thereof, and a method of use thereof |
US5770429A (en) | 1990-08-29 | 1998-06-23 | Genpharm International, Inc. | Transgenic non-human animals capable of producing heterologous antibodies |
US5122469A (en) | 1990-10-03 | 1992-06-16 | Genentech, Inc. | Method for culturing Chinese hamster ovary cells to improve production of recombinant proteins |
US5314695A (en) | 1990-11-13 | 1994-05-24 | Corvas International, Inc. | Tissue factor based prothrombin time reagent |
EP0564531B1 (en) | 1990-12-03 | 1998-03-25 | Genentech, Inc. | Enrichment method for variant proteins with altered binding properties |
US5248824A (en) * | 1990-12-31 | 1993-09-28 | The Biomembrane Institute | Method of preparing N,N,N-trimethylsphingosine |
US5137919A (en) * | 1990-12-31 | 1992-08-11 | Biomembrane Institute | Effect of N,N,N,-trimethylsphingosine on protein kinase C activity melanoma cell growth in vitro; metastatic potential in vivo and human platelet aggregation |
US5151360A (en) * | 1990-12-31 | 1992-09-29 | Biomembrane Institute | Effect of n,n,n-trimethylsphingosine on protein kinase-c activity, melanoma cell growth in vitro, metastatic potential in vivo and human platelet aggregation |
US5840867A (en) | 1991-02-21 | 1998-11-24 | Gilead Sciences, Inc. | Aptamer analogs specific for biomolecules |
US5677288A (en) * | 1991-05-15 | 1997-10-14 | Cypros Pharmaceutical Corporation | Use of aminoglycosides to protect against excitotoxic neuron damage |
DE69233482T2 (en) | 1991-05-17 | 2006-01-12 | Merck & Co., Inc. | Method for reducing the immunogenicity of antibody variable domains |
JP4124480B2 (en) | 1991-06-14 | 2008-07-23 | ジェネンテック・インコーポレーテッド | Immunoglobulin variants |
US5565332A (en) | 1991-09-23 | 1996-10-15 | Medical Research Council | Production of chimeric antibodies - a combinatorial approach |
US5430160A (en) | 1991-09-23 | 1995-07-04 | Florida State University | Preparation of substituted isoserine esters using β-lactams and metal or ammonium alkoxides |
US6025165A (en) | 1991-11-25 | 2000-02-15 | Enzon, Inc. | Methods for producing multivalent antigen-binding proteins |
ATE297465T1 (en) | 1991-11-25 | 2005-06-15 | Enzon Inc | METHOD FOR PRODUCING MULTIVALENT ANTIGEN-BINDING PROTEINS |
US5932448A (en) | 1991-11-29 | 1999-08-03 | Protein Design Labs., Inc. | Bispecific antibody heterodimers |
US5777085A (en) | 1991-12-20 | 1998-07-07 | Protein Design Labs, Inc. | Humanized antibodies reactive with GPIIB/IIIA |
WO1993016177A1 (en) | 1992-02-11 | 1993-08-19 | Cell Genesys, Inc. | Homogenotization of gene-targeting events |
US5714350A (en) | 1992-03-09 | 1998-02-03 | Protein Design Labs, Inc. | Increasing antibody affinity by altering glycosylation in the immunoglobulin variable region |
US6129914A (en) | 1992-03-27 | 2000-10-10 | Protein Design Labs, Inc. | Bispecific antibody effective to treat B-cell lymphoma and cell line |
US5573905A (en) | 1992-03-30 | 1996-11-12 | The Scripps Research Institute | Encoded combinatorial chemical libraries |
US5260288A (en) * | 1992-04-03 | 1993-11-09 | The Biomembrane Institute | Method for inhibition of cell motility by sphingosine-1-phosphate and derivatives |
US5369030A (en) * | 1992-09-11 | 1994-11-29 | Duke University | Method of inducing cellular differentiations and altering cell phenotype using ceramide analogs |
US5288514A (en) | 1992-09-14 | 1994-02-22 | The Regents Of The University Of California | Solid phase and combinatorial synthesis of benzodiazepine compounds on a solid support |
GB9223377D0 (en) | 1992-11-04 | 1992-12-23 | Medarex Inc | Humanized antibodies to fc receptors for immunoglobulin on human mononuclear phagocytes |
US5488038A (en) | 1992-11-27 | 1996-01-30 | Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai | Dibekacin derivatives and arbekacin derivatives active against resistant bacteria |
US6210671B1 (en) | 1992-12-01 | 2001-04-03 | Protein Design Labs, Inc. | Humanized antibodies reactive with L-selectin |
JP3852945B2 (en) | 1993-01-22 | 2006-12-06 | スローン − ケタリング・インスティテュート・フォー・キャンサー・リサーチ | Ganglioside-KLH complex with QS-21 |
CN1040177C (en) | 1993-04-23 | 1998-10-14 | 江苏省微生物研究所 | 1-N-ethyl gentamicin derivative and its preparing method |
US5444087A (en) * | 1993-10-19 | 1995-08-22 | Bristol-Myers Squibb Company | Manumycin compounds |
CA2176220A1 (en) * | 1993-11-15 | 1995-05-26 | Elizabeth Anne Woodcock | A method for treating cardiac dysfunction and pharmaceutical compositions useful therefor |
GB9325182D0 (en) | 1993-12-08 | 1994-02-09 | T Cell Sciences Inc | Humanized antibodies or binding proteins thereof specific for t cell subpopulations exhibiting select beta chain variable regions |
US5519134A (en) | 1994-01-11 | 1996-05-21 | Isis Pharmaceuticals, Inc. | Pyrrolidine-containing monomers and oligomers |
DE69518627T2 (en) | 1994-02-02 | 2001-01-11 | Liposome Co Inc | PHARMACEUTICALLY EFFECTIVE COMPOUNDS AND LIPOSOME AND METHOD FOR THE PRODUCTION THEREOF |
US5593853A (en) | 1994-02-09 | 1997-01-14 | Martek Corporation | Generation and screening of synthetic drug libraries |
US5585476A (en) * | 1994-02-15 | 1996-12-17 | Maclennan; Alexander J. | Molecular cloning and expression of G-protein coupled receptors |
US5539083A (en) | 1994-02-23 | 1996-07-23 | Isis Pharmaceuticals, Inc. | Peptide nucleic acid combinatorial libraries and improved methods of synthesis |
US5627171A (en) * | 1994-04-11 | 1997-05-06 | Oncomembrane, Inc. | Sphingosine-1-phosphate/trimethylsphingosine composition |
US5534615A (en) | 1994-04-25 | 1996-07-09 | Genentech, Inc. | Cardiac hypertrophy factor and uses therefor |
US5525735A (en) | 1994-06-22 | 1996-06-11 | Affymax Technologies Nv | Methods for synthesizing diverse collections of pyrrolidine compounds |
US5549974A (en) | 1994-06-23 | 1996-08-27 | Affymax Technologies Nv | Methods for the solid phase synthesis of thiazolidinones, metathiazanones, and derivatives thereof |
US6323201B1 (en) * | 1994-12-29 | 2001-11-27 | The Regents Of The University Of California | Compounds for inhibition of ceramide-mediated signal transduction |
US6046037A (en) | 1994-12-30 | 2000-04-04 | Hiatt; Andrew C. | Method for producing immunoglobulins containing protection proteins in plants and their use |
US5731168A (en) | 1995-03-01 | 1998-03-24 | Genentech, Inc. | Method for making heteromultimeric polypeptides |
US6096871A (en) | 1995-04-14 | 2000-08-01 | Genentech, Inc. | Polypeptides altered to contain an epitope from the Fc region of an IgG molecule for increased half-life |
US5702892A (en) | 1995-05-09 | 1997-12-30 | The United States Of America As Represented By The Department Of Health And Human Services | Phage-display of immunoglobulin heavy chain libraries |
US7060808B1 (en) | 1995-06-07 | 2006-06-13 | Imclone Systems Incorporated | Humanized anti-EGF receptor monoclonal antibody |
AU6113396A (en) | 1995-06-14 | 1997-01-15 | Regents Of The University Of California, The | Novel high affinity human antibodies to tumor antigens |
US5677189A (en) | 1995-06-29 | 1997-10-14 | Oncomembrane, Inc. | Method for quantifying sphingosine and for diagnosing platelet activation |
US5569588A (en) | 1995-08-09 | 1996-10-29 | The Regents Of The University Of California | Methods for drug screening |
US5667337A (en) * | 1995-09-15 | 1997-09-16 | Lazes; Richard J. | Rotating containment and repelling boom and method for confining a material floatable on a liquid surface |
WO1997010817A1 (en) * | 1995-09-20 | 1997-03-27 | The Regents Of The University Of Michigan | Amino ceramide-like compounds and therapeutic methods of use |
US6121246A (en) | 1995-10-20 | 2000-09-19 | St. Elizabeth's Medical Center Of Boston, Inc. | Method for treating ischemic tissue |
US6031071A (en) | 1996-01-24 | 2000-02-29 | Biophage, Inc. | Methods of generating novel peptides |
US5882644A (en) | 1996-03-22 | 1999-03-16 | Protein Design Labs, Inc. | Monoclonal antibodies specific for the platelet derived growth factor β receptor and methods of use thereof |
US5834597A (en) | 1996-05-20 | 1998-11-10 | Protein Design Labs, Inc. | Mutated nonactivating IgG2 domains and anti CD3 antibodies incorporating the same |
US5830916A (en) * | 1996-05-23 | 1998-11-03 | Duke University | Inhibitor of ceramidase |
DE19621038A1 (en) * | 1996-05-24 | 1997-11-27 | Boehringer Ingelheim Kg | Aminoguanidines, processes for their preparation and medicaments containing these compounds |
DE19636255C1 (en) | 1996-09-06 | 1997-10-23 | Fresenius Medical Care De Gmbh | High availability dialysis unit with automatic removal of accumulated lime-scale deposits |
US6013256A (en) | 1996-09-24 | 2000-01-11 | Protein Design Labs, Inc. | Method of preventing acute rejection following solid organ transplantation |
US6140060A (en) * | 1996-12-12 | 2000-10-31 | Chun; Jerold J. M. | Cloned lysophosphatidic acid receptors |
US5919687A (en) * | 1996-12-24 | 1999-07-06 | John Hopkins University | Recombinant N-SMases and nucleic acids encoding same |
US6310191B1 (en) | 1998-02-02 | 2001-10-30 | Cosmix Molecular Biologicals Gmbh | Generation of diversity in combinatorial libraries |
US5912144A (en) | 1997-04-24 | 1999-06-15 | Incyte Pharmaceuticals, Inc. | Edg-1-receptor homolog |
GB9710699D0 (en) | 1997-05-24 | 1997-07-16 | Danbiosyst Uk | Gastro-retentive controlled release system |
CA2293718A1 (en) * | 1997-06-10 | 1998-12-17 | Medlyte Diagnostics, Inc. | Methods for early detection of heart disease |
US5989803A (en) | 1997-09-05 | 1999-11-23 | The Trustees Of Columbia University In The City Of New York | Method for treating a subject suffering from a condition associated with an extracellular zinc sphingomyelinase |
US6649362B2 (en) | 1997-09-08 | 2003-11-18 | Medvet Science Pty. Ltd. | Screening method for an agent having an effect on a sphingosine kinase signaling pathway |
US6423527B1 (en) * | 1997-09-29 | 2002-07-23 | Children's Hospital Medical Center Of Northern California | Sphingosine-1-phosphate lyase polypeptides, polynucleotides and modulating agents and methods of use therefor |
BR9813365A (en) | 1997-12-05 | 2004-06-15 | Scripps Research Inst | Method for Production and Humanization of a Mouse Monoclonal Antibody |
US6057126A (en) * | 1997-12-24 | 2000-05-02 | Allelix Biopharmaceuticals, Inc. | Mammalian EDG-5 receptor homologs |
US6098631A (en) | 1998-01-21 | 2000-08-08 | The Regents Of The University Of Michigan | Compositions and methods for treating autoimmune disease |
WO1999038983A1 (en) * | 1998-01-29 | 1999-08-05 | Smithkline Beecham Plc | Novel sphingosine-1 phosphate lyase |
WO1999041266A1 (en) | 1998-02-12 | 1999-08-19 | Emory University | Sphingolipid derivatives and their methods of use |
WO1999045959A1 (en) | 1998-03-13 | 1999-09-16 | Dana-Farber Cancer Institute, Inc. | Humanized antibody and uses thereof |
US6130067A (en) * | 1998-05-20 | 2000-10-10 | Smithkline Beecham Corporation | Human EDG3sb gene |
US6130235A (en) | 1998-05-22 | 2000-10-10 | Scios Inc. | Compounds and methods to treat cardiac failure and other disorders |
DE69939651D1 (en) * | 1998-06-29 | 2008-11-13 | Los Angeles Childrens Hospital | TREATMENT OF HYPERPROLIFERATIVE DISEASES |
DE69942671D1 (en) | 1998-12-01 | 2010-09-23 | Facet Biotech Corp | HUMANIZED ANTIKOERPER AGAINST GAMMA INTERFERON |
WO2001037836A1 (en) | 1999-11-24 | 2001-05-31 | Emory University | Diimino-piperazine derivatives for use as modulators of cell regulation |
US6306911B1 (en) * | 2000-02-07 | 2001-10-23 | Ortho-Mcneil Pharmaceutical, Inc. | Substituted amino acids as neutral sphingomyelinase inhibitors |
JP2001261575A (en) * | 2000-03-13 | 2001-09-26 | General Hospital Corp | Method for regulating vasoconstriction and its composition |
US6571638B2 (en) | 2000-06-30 | 2003-06-03 | Sawtek, Inc. | Surface-acoustic-wave pressure sensor and associated methods |
AU2001288563A1 (en) | 2000-08-31 | 2002-03-13 | University Of Connecticut | Regulation of angiogenesis via modulation of edg receptor mediated signal transduction comprising sphingosine-1-phosphate administration |
US20030125533A1 (en) * | 2000-10-06 | 2003-07-03 | Sophia Kossida | Regulation of human sphingosine kinase-like protein |
EP1363643A2 (en) | 2000-12-22 | 2003-11-26 | Medlyte, Inc. | Compositions and methods for the treatment and prevention of cardiovascular diseases and disorders, and for identifying agents therapeutic therefor |
US20020150582A1 (en) * | 2001-02-08 | 2002-10-17 | Friedrichs Gregory S. | Method of treating or inhibiting cellular injury or cell death |
JP2002243737A (en) * | 2001-02-09 | 2002-08-28 | Azwell Inc | Anti-sphingolipid monoclonal antibody |
US6868383B1 (en) * | 2001-07-12 | 2005-03-15 | At&T Corp. | Systems and methods for extracting meaning from multimodal inputs using finite-state devices |
US7674580B2 (en) * | 2002-01-17 | 2010-03-09 | Children's Hospital & Research Center At Oakland | Compositions and methods for the modulation of sphingolipid metabolism and/or signaling |
KR20120125398A (en) | 2002-05-16 | 2012-11-14 | 노파르티스 아게 | Use of edg receptor binding agents in cancer |
US7794713B2 (en) * | 2004-04-07 | 2010-09-14 | Lpath, Inc. | Compositions and methods for the treatment and prevention of hyperproliferative diseases |
DK1812797T3 (en) * | 2004-10-28 | 2013-05-21 | Lpath Inc | Compositions and Methods for the Treatment and Prevention of Hyperproliferative Diseases |
WO2006104989A2 (en) | 2005-03-29 | 2006-10-05 | Verenium Corporation | Altered antibody fc regions and uses thereof |
-
2005
- 2005-10-28 US US11/261,935 patent/US7794713B2/en not_active Expired - Fee Related
-
2006
- 2006-10-27 SI SI200631487T patent/SI1948234T1/en unknown
- 2006-10-27 PL PL06844226T patent/PL1948234T3/en unknown
- 2006-10-27 AU AU2006309008A patent/AU2006309008B2/en not_active Ceased
- 2006-10-27 US US12/091,890 patent/US20090220523A1/en not_active Abandoned
- 2006-10-27 WO PCT/US2006/042027 patent/WO2007053447A2/en active Application Filing
- 2006-10-27 CA CA002627427A patent/CA2627427A1/en not_active Abandoned
- 2006-10-27 EP EP06844226A patent/EP1948234B1/en not_active Not-in-force
- 2006-10-27 DK DK06844226.8T patent/DK1948234T3/en active
- 2006-10-27 EA EA200801197A patent/EA017945B1/en not_active IP Right Cessation
- 2006-10-27 CN CNA2006800483373A patent/CN101340929A/en active Pending
- 2006-10-27 PT PT68442268T patent/PT1948234E/en unknown
- 2006-10-27 ES ES06844226T patent/ES2398919T3/en active Active
- 2006-10-27 JP JP2008538001A patent/JP5739089B2/en not_active Expired - Fee Related
- 2006-10-27 US US11/588,973 patent/US20070148168A1/en not_active Abandoned
-
2009
- 2009-01-23 HK HK09100789.6A patent/HK1123000A1/en not_active IP Right Cessation
-
2014
- 2014-09-01 JP JP2014176894A patent/JP2015057382A/en active Pending
Non-Patent Citations (1)
Title |
---|
See references of EP1948234A4 * |
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US8026342B2 (en) * | 2006-10-27 | 2011-09-27 | Lpath, Inc. | Compositions and methods for binding sphingosine-1-phosphate |
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US9163091B2 (en) | 2007-05-30 | 2015-10-20 | Lpath, Inc. | Compositions and methods for binding lysophosphatidic acid |
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WO2008150841A1 (en) * | 2007-05-30 | 2008-12-11 | Lpath, Inc. | Compositions and methods for binding lysophosphatidic acid |
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Also Published As
Publication number | Publication date |
---|---|
PL1948234T3 (en) | 2013-06-28 |
US20060171946A1 (en) | 2006-08-03 |
SI1948234T1 (en) | 2013-02-28 |
US20090220523A1 (en) | 2009-09-03 |
JP5739089B2 (en) | 2015-06-24 |
WO2007053447A3 (en) | 2007-07-05 |
AU2006309008A1 (en) | 2007-05-10 |
EA200801197A1 (en) | 2009-02-27 |
CN101340929A (en) | 2009-01-07 |
CA2627427A1 (en) | 2007-05-10 |
HK1123000A1 (en) | 2009-06-05 |
EP1948234B1 (en) | 2012-12-05 |
ES2398919T3 (en) | 2013-03-22 |
JP2015057382A (en) | 2015-03-26 |
DK1948234T3 (en) | 2013-01-28 |
AU2006309008B2 (en) | 2013-06-20 |
US20070148168A1 (en) | 2007-06-28 |
EP1948234A4 (en) | 2010-06-09 |
EP1948234A2 (en) | 2008-07-30 |
JP2009513668A (en) | 2009-04-02 |
US7794713B2 (en) | 2010-09-14 |
EA017945B1 (en) | 2013-04-30 |
PT1948234E (en) | 2013-02-11 |
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