Viral Conjunctivitis Treatment Using Ranpirnase and/or Amphinase

a technology of ranpirin and amphinase, which is applied in the field of viral conjunctivitis treatment using ranpirin and/or amphinase, can solve the problems of high contagiousness of bacteria and the lack of effective treatment for viral conjunctivitis, and achieve the effects of reducing a symptom, and reducing or suppressing a level of virus

Inactive Publication Date: 2017-09-14
OKOGEN INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0006]Aspects of the present specification disclose a method of treating an individual with a viral conjunctivitis. The disclosed method comprises administering to an individual in need thereof a pharmaceutical composition comprising a therapeutic effective amount of one or more Ranpirnases and / or a therapeutic effective amount of one or more Amphinase. Such administration reduces a symptom associated with the viral conjunctivitis, thereby treating the individual. Also disclosed is a use of a pharmaceutical composition comprising one or more Ranpirnases and / or one or more Amphinase for treating of a viral conjunctivitis. A viral conjunctivitis disclosed herein includes an epidemic keratoconjunctivitis, a pharyngoconjunctival fever, a nonspecific sporadic follicular conjunctivitis, or a chronic papillary conjunctivitis. A viral conjunctivitis disclosed herein may be caused by a Human adenovirus B, a Human adenovirus D, or a Human adenovirus E.
[0007]Other aspects of methods of reducing or suppressing a level of a virus, viral titer, viral replication, protein synthesis and / or tRNA in an individual. The disclosed methods comprises administering to an individual in need thereof a pharmaceutical composition comprising a therapeutic effective amount of one or more Ranpirnases and / or a therapeutic effective amount of one or more Amphinase. Such administration reduces or suppresses a level of a virus, viral titer, viral replication, protein synthesis and / or tRNA. Also disclosed is a use of a pharmaceutical composition comprising one or more Ranpirnases and / or one or more Amphinase for reducing or suppressing a level of a virus, viral titer, viral replication, protein synthesis and / or tRNA in an individual.
[0008]Other aspects of a method of reducing or suppressing a level of an inflammation inducing molecule in an individual. The disclosed method comprises administering to an individual in need thereof a pharmaceutical composition comprising a therapeutic effective amount of one or more Ranpirnases and / or a therapeutic effective amount of one or more Amphinase. Such administration reduces or suppresses a level of an inflammation inducing molecule. Also disclosed is a use of a pharmaceutical composition comprising one or more Ranpirnases and / or one or more Amphinase for reducing or suppressing a level of an inflammation inducing molecule. An inflammation inducing molecule disclosed herein includes a substance P, a calcitonin gene-related peptide, a glutamate, or a combination thereof.
[0009]Other aspects of a method of reducing or suppressing a level of an inflammation inducing prostaglandin in an individual. The disclosed method comprises administering to an individual in need thereof a pharmaceutical composition comprising a therapeutic effective amount of one or more Ranpirnases and / or a therapeutic effective amount of one or more Amphinase. Such administration reduces or suppresses a level of an inflammation inducing prostaglandin. Also disclosed is a use of a pharmaceutical composition comprising one or more Ranpirnases and / or one or more Amphinase for reducing or suppressing a level of an inflammation inducing prostaglandin. An inflammation inducing prostaglandin includes 15dPGJ2.
[0010]Other aspects of a method of stimulating or enhancing a peroxisome proliferator-activated receptor (PPAR) signaling pathway activity in an individual. The disclosed method comprises administering to an individual in need thereof a pharmaceutical composition comprising a therapeutic effective amount of one or more Ranpirnases and / or a therapeutic effective amount of one or more Amphinase. Such administration stimulates or enhances a PPAR signaling pathway activity. Also disclosed is a use of a pharmaceutical composition comprising one or more Ranpirnases and / or one or more Amphinase for stimulating or enhancing a PPAR signaling pathway activity. A PPAR signaling pathway activity includes a PPAR-α signaling pathway activity, a PPAR-γ signaling pathway activity, and a PPAR-δ (also known as PPAR-β) signaling pathway activity
[0011]Other aspects of a method of promoting the resolving phenotypic change of M1 to M2 in an individual. The disclosed method comprises administering to an individual in need thereof a pharmaceutical composition comprising a therapeutic effective amount of one or more Ranpirnases and / or a therapeutic effective amount of one or more Amphinase. Such administration induces apoptosis of Macrophage M1 cells, promotes differentiation of Macrophage M2 cells or both, thereby promoting the resolving phenotypic change of M1 to M2. Also disclosed is a use of a pharmaceutical composition comprising one or more Ranpirnases and / or one or more Amphinase for promoting the resolving phenotypic change of M1 to M2.

Problems solved by technology

Both viral and bacterial conjunctivitis are highly contagious.
Unlike bacterial infection, there is currently no effective treatment for viral conjunctivitis.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

In Vitro Antiviral Effects of Ranpirnase

[0215]The antiviral activity of Ranpirnase was evaluated using three in vitro assays. In each assay, Human Adenovirus 5 (AD5 strain NYS #98-1836) or Human Adenovirus 8 (Ad8 strain 13306) was used to infect cells in order to determine whether Ranpirnase could inhibited the cytopathic effects of virus in infected cells and / or could reduce the amount of virus being produced from those infected cells.

[0216]To assess in vitro cytotoxicity, both a cytopathic effects of a virus (CPE) regression assay and a neutral red lysosomal uptake cell viability assay was performed essentially as described in, e.g., Repetto, et al., Neutral Red Uptake Assay for the Estimation of Cell Viability / Cytotoxicity, Nat. Protoc. 3: 1125-1131 (2008), the content of which is hereby incorporated by reference in its entirety. Ninety-six-well plates were seeded with MA-104 cells at 1×105 cells / mL and incubated overnight at 37° C. with 5% CO2. Ranpirnase was serially diluted in...

example 2

Cytotoxicity Effects of Ranpirnase

[0221]To confirm the cytotoxicity effects of Ranpirnase a plaque reduction assay was performed essentially as described in, e.g., Romanowski, et al., The In Vitro and In Vivo Evaluation of ddC as a Topical Antiviral for Ocular Adenovirus Infections, Invest. Ophthalmol. Vis. Sci. 50: 5295-5299 (2009), which is hereby incorporated by reference in its entirety.

[0222]To assess in vitro cytotoxicity, 96-well plates were seeded with A549 cells at 1×105 cells / mL and incubated overnight at 37° C. with 5% CO2. Ranpirnase was serially diluted to concentrations of 1.0 μM, 10 μM and 50 μM. After removal of the tissue culture media, 100 μL of each dilution was added to 3 wells of a 96-well plate with 80% to 100% confluent cells. As controls, 100 μL of a lysis buffer containing 0.25% TRITON X-100 was added to 6 wells (positive cytotoxicity control) and 100 μL of tissue culture media with no Ranpirnase was added to 6 wells (negative cytotoxicity control). Each tes...

example 3

In Vivo Antiviral Effects of Ranpirnase

[0226]It spite of its poor cytotoxicity profile, the antiviral activity of Ranpirnase was evaluated in vivo using an ocular rabbit replication model. See, e.g., Romanowski, et al., The In Vitro and In Vivo Evaluation of ddC as a Topical Antiviral for Ocular Adenovirus Infections, Invest. Ophthalmol. Vis. Sci. 50: 5295-5299 (2009), which is hereby incorporated by reference in its entirety.

[0227]To conduct this in vivo assay, 25 NZW rabbit were anesthetized using the general anesthesia ketamine and xylazine and the topical anesthesia proparacaine. Each rabbit was then topically inoculated with 50 μL of Adenovirus serotype Ad5 (3×107 pfu / mL) in both eyes after corneal epithelial scarification (12 cross-hatched strokes of a 25 sterile needle). Eyes were closed and gently rubbed for 5 seconds to ensure contact of the virus on all ocular surfaces. Inoculation of both eyes allowed for the reduction in the number of animals needed without jeopardizing ...

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Abstract

The present specification discloses Ranpirnase and Amphinase, compositions comprising Ranpirnase and/or Amphinase, and methods and uses to treat a viral conjunctivitis, an epidemic keratoconjunctivitis, and/or a pharyngoconjunctival fever, reduce or suppress a level of virus or viral titer, reduce or suppress viral replication, reduce or suppress protein synthesis, reduce or suppress a level of a tRNA, reduce or suppress a level of an inflammation inducing molecule and/or an inflammation inducing prostaglandin, stimulate or enhance a peroxisome proliferator-activated receptor (PPAR) pathway signal, promote the resolving phenotypic change of M1 to M2, modulate Th1 and Th2 cytokines, and/or reduce or suppress a NFκB pathway signal using Ranpirnase, Amphinase or compositions comprising Ranpirnase and/or Amphinase.

Description

[0001]This continuation application claims the benefit of priority and the filing date pursuant to 35 U.S.C. §120 of U.S. Non-Provisional patent application Ser. No. 15 / 275,442, filed on Sep. 25, 2016, an application which claims priority and the filing date pursuant 35 U.S.C. §119(e) of U.S. Provisional Patent Application 62 / 233,267, filed on Sep. 25, 2015, the content of each of which is hereby incorporated by reference in its entirety.[0002]Conjunctivitis, commonly referred to as pink eye, is an inflammation of the eye causing swelling and irritation. It affects the conjunctiva, the thin transparent membrane that covers the sclera of the eyeball and lines the inner surface of the eyelid. Conjunctivitis is most often caused by a viral or by a bacterial infection, although allergies, chemical irritants, and underlying diseases can also play a role. Symptoms of conjunctivitis include, without limitation, redness in the sclera and / or inner eyelid, ocular itching (itchy eyes), foreign...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/46A61K9/00
CPCA61K38/465C12Y301/27A61K9/0048A61K9/0051C12Y301/27005A61P27/02C12N15/113
Inventor STREM, BRIAN
Owner OKOGEN INC
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