TREATMENTS FOR DISEASES AND DISORDERS THAT INVOLVE OXIDATIVE STRESS

Abstract

Peptide compounds and their use in treating diseases and disorders that cause, are caused by, or are characterized by cellular oxidative stress.

Claims

1. A compound comprising or consisting of an amino acid sequence having the formula:
Gly-X-Thr-Pro wherein X is a three amino acid sequence selected from: Arg-Ala-Cys(Acid); Arg-Gly-Cys; Arg-Asp-Gly; Arg-Ala-Glu; Arg-Gly-Asn; Asp-Gly-Arg; Cys(acid)-Gly-Arg and Lys-Gly-Asp.

2. A compound comprising or consisting of the amino acid sequence Gly-Arg-Gly-Cys(acid)-Gly-Gly-Gly-Asp-Gly.

3. A compound according to claim 1 wherein one or more additional amino acids are added to either end or both ends of the compound such that the compound consists of more than six, but no more than ten, amino acids.

4. A compound according to claim 1 wherein the N-terminal Gly is replaced by another amino acid or group selected from: Arg, Asp, His, Lys, Trp, Phe, Tyr, Met, Ala, Leu, Guanidino.

5. A compound according to claim 1 wherein one or both of Thr-Pro at the C-terminal end is/are replaced by one or two other amino acids selected from: Ser, Val, Thr, Phe, Tyr, Lys, Asp.

6. A compound according to claim 1 prepared as a salt.

7. A compound according to claim 6 comprising a salt form selected from: hydrochloride, acetate, trifluoroacetate.

8. A pharmaceutical preparation comprising a compound according to claim 1 in combination with at least one pharmaceutically acceptable carrier, diluent, solvent or excipient.

9-23. (canceled)

24. A compound according to claim 1 wherein the compound comprises ALG-3001.

25. A compound according to claim 1, wherein the compound comprises ALG-3002.

26. A compound according to claim 1, wherein the compound comprises ALG-3003.

27. A compound according to claim 1, wherein the compound comprises ALG-3004.

28. A compound according to claim 1, wherein the compound comprises ALG-3005.

29. A compound according to claim 2, wherein the compound comprises ALG-3006.

30. A compound according to claim 1, wherein the compound comprises ALG-3007.

31. A compound according to claim 1, wherein the compound comprises ALG-3008.

32. A compound according to claim 1, wherein the compound comprises ALG-3009.

33-36. (canceled)

37. A pharmaceutical preparation comprising a compound according to claim 1 for intravitreal injection or implantation.

38. A pharmaceutical preparation comprising a compound according to claim 1 for topical administration to the eye.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0028] FIG. 1A is a bar graph comparing the effects of GRGETP (Control Peptide), Risuteganib (RSG) (Positive Control) and ALG-3001 (P1) on retinal neovascularization in OIR (ocular ischemic retinopathy) mice in a first experiment.

[0029] FIG. 1B is a bar graph comparing the effects of GRGETP (Control Peptide), Risuteganib (RSG) (Positive Control) and ALG-3001 (P1) on retinal neovascularization in OIR mice in a second experiment.

[0030] FIG. 1C is a bar graph comparing the effects of Control (serum-free, phenol red-free MEM (SF-MEM)), 150 ?M Hydroquinone (HQ) alone, and 150 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3001 (P1), 400 ?M ALG-3001 (P1) and 800 ?M ALG-3001 (P1). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, *P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001)

[0031] FIG. 1D is a bar graph comparing the effects of Control (SF-MEM), 170 ?M Hydroquinone (HQ) alone, and 170 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3001 (P1), 400 ?M ALG-3001 (P1) and 800 ?M ALG-3001 (P1). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001) FIG. 2 is a bar graph comparing the effects of GRGETP (Control Peptide), Risuteganib (RSG) (Positive Control) and ALG-3002 (P2) on retinal neovascularization in OIR mice.

[0032] FIG. 2 is a bar graph comparing the effects of GRGETP (Control Peptide), Risuteganib (RSG) (Positive Control) and ALG-3002 (P2) on retinal neovascularization in OIR mice in a second experiment.

[0033] FIG. 2A is a bar graph comparing the effects of Control (SF-MEM), 170 ?M Hydroquinone (HQ) alone, and 170 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3002 (P2), 400 ?M ALG-3002 (P2) and 800 ?M ALG-3002 (P2). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, *P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0034] FIG. 2B is a bar graph comparing the effects of Control (SF-MEM), 160 ?M Hydroquinone (HQ) alone, and 160 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3002 (P2), 400 ?M ALG-3002 (P2) and 800 ?M ALG-3002 (P2). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, *P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0035] FIG. 2C is a bar graph comparing the effects of Control (SF-MEM), 140 ?M Hydroquinone (HQ) alone, and 140 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 12.5 ?M ALG-3002 (P2), 25 ?M ALG-3002 (P2), 50 ?M ALG-3002 (P2) and 100 ?M ALG-3002 (P2).

[0036] FIG. 2D is a bar graph comparing the effects of Control (SF-MEM), 160 ?M Hydroquinone (HQ) alone, and 160 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 12.5 ?M ALG-3002 (P2), 25 ?M ALG-3002 (P2), 50 ?M ALG-3002 (P2) and 100 ?M ALG-3002 (P2).

[0037] FIG. 3 is a bar graph comparing the effects of GRGETP (Control Peptide) and ALG-3003 (P3) on retinal neovascularization in OIR mice.

[0038] FIG. 3A is a bar graph comparing the effects of Control (SF-MEM), 140 ?M Hydroquinone (HQ) alone, and 140 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3003 (P3), 400 ?M ALG-3003 (P3) and 800 ?M ALG-3003 (P3). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, *P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001)

[0039] FIG. 3B is a bar graph comparing the effects of Control (SF-MEM), 160 ?M Hydroquinone (HQ) alone, and 160 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3003 (P3), 400 ?M ALG-3003 (P3) and 800 ?M ALG-3003 (P3). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, *P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0040] FIG. 4 is a bar graph comparing the effects of GRGETP (Control Peptide) and ALG-3004 (P4) on retinal neovascularization in OIR mice.

[0041] FIG. 4A is a bar graph comparing the effects of Control (SF-MEM), 140 ?M Hydroquinone (HQ) alone, and 140 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3004 (P4), 400 ?M ALG-3004 (P4) and 800 ?M ALG-3004 (P4). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0042] FIG. 4B is a bar graph comparing the effects of Control (SF-MEM), 160 ?M Hydroquinone (HQ) alone, and 160 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3004 (P4), 400 ?M ALG-3004 (P4) and 800 ?M ALG-3004 (P4). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0043] FIG. 5 is a bar graph comparing the effects of GRGETP (Control Peptide) and ALG-3005 (P5) on retinal neovascularization in OIR mice.

[0044] FIG. 5A is a bar graph comparing the effects of Control (SF-MEM), 140 ?M Hydroquinone (HQ) alone, and 140 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3005 (P5), 400 ?M ALG-3005 (P5) and 800 ?M ALG-3005 (P5). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0045] FIG. 5B is a bar graph comparing the effects of Control (SF-MEM), 160 ?M Hydroquinone (HQ) alone, and 160 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3005 (P5), 400 ?M ALG-3005 (P5) and 800 ?M ALG-3005 (P5). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0046] FIG. 6 is a bar graph comparing the effects of GRGETP (Control Peptide) and ALG-3006 (P6) on retinal neovascularization in OIR mice.

[0047] FIG. 6A is a bar graph comparing the effects of Control (SF-MEM), 140 ?M Hydroquinone (HQ) alone, and 140 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3006 (P6), 400 ?M ALG-3006 (P6) and 800 ?M ALG-3006 (P6). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0048] FIG. 6B is a bar graph comparing the effects of Control (SF-MEM), 160 ?M Hydroquinone (HQ) alone, and 160 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3006 (P6), 400 ?M ALG-3006 (P6) and 800 ?M ALG-3006 (P6). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0049] FIG. 7 is a bar graph comparing the effects of GRGETP (Control Peptide) and ALG-3007 (P7) on retinal neovascularization in OIR mice.

[0050] FIG. 7A is a bar graph comparing the effects of Control (SF-MEM), 140 ?M Hydroquinone (HQ) alone, and 140 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3007 (P7), 400 ?M ALG-3007 (P7) and 800 ?M ALG-3007 (P7). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0051] FIG. 7B is a bar graph comparing the effects of Control (SF-MEM), 160 ?M Hydroquinone (HQ) alone, and 160 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3007 (P7), 400 ?M ALG-3007 (P7) and 800 ?M ALG-3007 (P7). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0052] FIG. 8 is a bar graph comparing the effects of GRGETP (Control Peptide), Risuteganib (RSG) (Positive Control) and ALG-3008 (P8) on retinal neovascularization in OIR mice.

[0053] FIG. 8A is a bar graph comparing the effects of Control (SF-MEM), 140 ?M Hydroquinone (HQ) alone, and 140 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3008 (P8), 400 ?M ALG-3008 (P8) and 800 ?M ALG-3008 (P8). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, *P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0054] FIG. 8B is a bar graph comparing the effects of Control (SF-MEM), 160 ?M Hydroquinone (HQ) alone, and 160 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3008 (P8), 400 ?M ALG-3008 (P8) and 800 ?M ALG-3008 (P8). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0055] FIG. 9 is a bar graph comparing the effects of GRGETP (Control Peptide) and ALG-3009 (P9) on retinal neovascularization in OIR mice.

[0056] FIG. 9A is a bar graph comparing the effects of Control (SF-MEM), 140 ?M Hydroquinone (HQ) alone, and 140 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3009 (P9), 400 ?M ALG-3009 (P9) and 800 ?M ALG-3009 (P9). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0057] FIG. 9B is a bar graph comparing the effects of Control (SF-MEM), 160 ?M Hydroquinone (HQ) alone, and 160 ?M Hydroquinone (HQ) in combination with each of: 400 ?M Risuteganib (RSG), 100 ?M ALG-3009 (P9), 400 ?M ALG-3009 (P9) and 800 ?M ALG-3009 (P9). Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0058] FIG. 10 is a bar graph showing the effects of ALG-3002 (P2) on cell viability in: normal (unstressed) retinal pigment epithelium (RPE) cells, Hydroquinone(HQ)-stressed retinal pigment epithelium (RPE) cells, and carbonyl cyanide phenylhydrazone (CCCP)-stressed retinal pigment epithelium (RPE) cells. Statistical significance or non-significance is denoted as follows: (N.S.=not significant, *P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0059] FIG. 11 is a bar graph showing the effects of ALG-3002 (P2) on Reactive Oxygen Species (ROS) in: normal (unstressed) retinal pigment epithelium (RPE) cells and Hydroquinone(HQ)-stressed retinal pigment epithelium (RPE) cells. Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0060] FIG. 12 is a bar graph showing the effects of ALG-3004 (P4) on Reactive Oxygen Species (ROS) in: normal (unstressed) retinal pigment epithelium (RPE) cells and Hydroquinone(HQ)-stressed retinal pigment epithelium (RPE) cells. Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0061] FIG. 13 is a bar graph showing the effects of ALG-3007 (P7) on Reactive Oxygen Species (ROS) in: normal (unstressed) retinal pigment epithelium (RPE) cells and Hydroquinone(HQ)-stressed retinal pigment epithelium (RPE) cells. Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0062] FIG. 14 is a bar graph showing the effects of ALG-3002 (P2) on Mitochondrial Membrane Potential (MMP) in: normal (unstressed) retinal pigment epithelium (RPE) cells and Hydroquinone(HQ)-stressed retinal pigment epithelium (RPE) cells. Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0063] FIG. 15 is a bar graph showing the effects of ALG-3004 (P4) on Mitochondrial Membrane Potential (MMP) in: normal (unstressed) retinal pigment epithelium (RPE) cells and Hydroquinone(HQ)-stressed retinal pigment epithelium (RPE) cells. Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0064] FIG. 16 is a bar graph showing the effects of ALG-3007 (P7) on Mitochondrial Membrane Potential (MMP) in: normal (unstressed) retinal pigment epithelium (RPE) cells and Hydroquinone(HQ)-stressed retinal pigment epithelium (RPE) cells. Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0065] FIG. 17 is a bar graph comparing the effects of SF-MEM (Control), risuteganib (RSG) alone, ALG-3002 (P2) alone, hydroquinone (HQ) alone, hydroquinone (HQ)+risuteganib (RSG) and hydroquinone (HQ)+ALG-3002 (P2), on glutathione (GSH) concentration in retinal pigment epithelium (RPE) cells. Statistical significance or non-significance is denoted as follows: (N.S.=not significant, * P?0.05, ** P?0.01, *** P?0.001, and **** P?0.0001).

[0066] FIG. 18A shows micrographic images of mitochondrial morphology in normal MEM cultured retinal pigment epithelium (RPE) cells.

[0067] FIG. 18B shows micrographic images of mitochondrial morphology in hydroquinone (HQ)-stressed retinal pigment epithelium (RPE).

[0068] FIG. 18C shows micrographic images of mitochondrial morphology in hydroquinone (HQ)-stressed retinal pigment epithelium (RPE) cells that were treated with risuteganib (RSG).

[0069] FIG. 18D shows micrographic images of mitochondrial morphology in hydroquinone (HQ)-stressed retinal pigment epithelium (RPE) cells that were treated with ALG-3002 (P2).

DETAILED DESCRIPTION

[0070] The following detailed description and the accompanying drawings to which it refers are intended to describe some, but not necessarily all, examples or embodiments of the disclosed subject matter. The described examples or embodiments are to be considered in all respects only as illustrative and not restrictive. The contents of this detailed description and the accompanying drawings do not limit the scope of the disclosure in any way.

[0071] The treatments described in this patent application may be administered by any suitable route(s) of administration and in any suitable dosage form. Possible routes of administration known in the art include, but are not necessarily limited to: systemic, local, regional, parenteral, enteral, inhalational, topical, intramuscular, subcutaneous, intravenous, intravitreal, intra-arterial, intrathecal, intravesical, oral, endoscopic (e.g, through an endoscope, bronchoscope, colonoscope, sigmoidoscope, hysterscope, laproscope, athroscope, gastroscope, cystoscope, etc.), transurethral, trans-tympanic, rectal, nasal, oral, tracheal, bronchial, esophageal, gastric, intestinal, peritoneal, urethral, vesicular, urethral, vaginal, uterine, fallopian, buccal, lingual, sublingual and mucosal. Possible dosage forms known in the art include, but are not limited to: liquids, biphasic liquids, solids, semisolids, vapors, aerosols, solutions, suspensions, mixtures, syrups, linctuses, gels, creams, pastes, ointments, lotions, liniments, collodions, emulsions, transdermal delivery patches, suppositories, capsules, tablets, powders, granules, edibles, chewables, drops, sprays, enemas, douches, lozenges, etc.

In Vivo OIR/ROP Mouse Model Study of Retinal Neovascularization

[0072] In this study, Risuteganib (RSG), a known-active positive control having the amino acid sequence Gly-Arg-Gly-Cys(acid)-Thr-Pro, and each of Test Compounds (ALG-3001 through ALG-3009, alternately referred to as P1 through P9) were tested in comparison to a known inactive Control Peptide having the amino acid sequence Gly-Arg-Gly-Glu-Tyr-Pro (RGE) (e.g., a negative control) for effectiveness in reducing retinal neovascularization in a mouse model of oxygen-induced retinopathy (OIR).

[0073] Methods: OIR mouse pups received 5 days of hyperoxia (75% O2) to obliterate developing retinal vessels. Following their return to room air, retinal neovascularization develops due to an imbalance in oxygen supply and demand. At the time of return to room air, eyes of OIR pups received a single intravitreal injection of either Control Peptide (known inactive), Positive Control (known active) or Test Compound, as follows:

TABLE-US-00004 OIR STUDY TREATMENTS TREATMENT DESCRIPTION DOSE Control Peptide GRGETP 10 ?g per Eye Positive Control Risuteganib 10 ?g per Eye Test Compounds ALG-3001-ALG-3009 10 ?g per Eye

[0074] Injectate solutions were prepared by dissolving either Control Peptide, Risuteganib or a Test Compound in 0.9% NaCl saline at a concentration of 10 ?g per ?L. One microliter (1.0 ?L) of each solution was injected intraviterally into each eye, thereby delivering the indicated dose of 10 ?g per eye. Eighteen (18) days after injection, the mice were euthanized and the retinas were removed and stained with flurescein-labeled dextran for fluorescein microscopy. The prepared retinas were then examined microscopically and the area of each retina exhibiting neovascularization was measured.

[0075] Results: The results of this study are summarized in Table 3 (above), Table 4 (below) and in FIGS. 1A, 1B, 2, 3, 4, 5, 7, 8, 9 and 10.

TABLE-US-00005 TABLE 4 % NV Reduction Compared to Test Compound Control Peptide ALG-3001 (P1) 60 ALG-3001 (P1) 50 ALG-3002 (P2) 66 ALG-3003 (P3) 44 ALG-3004 (P4) 63 ALG-3005 (P5) 64 ALG-3006 (P6) 62 ALG-3007 (P7) 56 ALG-3008 (P8) 63 ALG-3009 (P9) 40

[0076] Risuteganib (RSG), the positive control, caused 47-64% reduction in neovascular area compared to the inactive Control Peptide (GRGETP). All Test Compounds (ALG-3001 (P1) through ALG-3009 (P9) caused a reduction in neovascular area comparable to that caused by risuteganib (RSG).

In vitro Cell Culture Study of ALG-3001 to ALG-3009 (P1 through P9) in Stressed Retinal Pigment Epithelium (RPE) Cells

[0077] In this study, ALG-3001 through ALG-3009 (alternately referred to as P1 through P9) were tested for their cytoprotective, mitochondrial stabilization and other therapeutic properties in a human RPE cell culture model. The cells were stressed with the cigarette smoke toxin, hydroquinone, which is known to reduce cell viability, elevate reactive oxygen species (ROS) and reduce mitochondrial function. This model replicates the disease scenario of retinal degenerations and specifically dry macular degeneration.

[0078] RPE cells form a monolayer of highly specialized, polarized epithelial cells interposed between the choriocapillaris and photoreceptors. RPE cells play an important role in retinal homeostasis and are vital to photoreceptor cell health and visual function.

[0079] RPE cell dysfunction or death is thought to be an important contributor to age-related macular degeneration (AMD). RPE cells are continually exposed to oxidants throughout life and oxidative stress plays a major role in AMD pathogenesis and progression. Cigarette smoke contains high concentrations of free oxidants and has been implicated as a major environmental risk factor for AMD. Hydroquinone (HQ), a major oxidant in both tobacco smoke and atmospheric pollutants, increases reactive oxygen species (ROS) generation and promotes oxidative stress.5 ROS, a group of unstable oxygen-containing molecules that can easily react with other molecules in a cell, are generated during cellular metabolism and in response to various stimuli. In cells, the major site of ROS production is the mitochondrial electron transport chain, where some electrons leak from the transport process and spontaneously react with molecular oxygen, producing superoxide anion. ROS have important physiological functions; however, excess ROS can cause RPE cell oxidative damage.

[0080] Methods: Human donor eyes from a 62-year-old male donor were obtained from an Organ Donor and Eye Bank in accordance with the provisions of the Declaration of Helsinki for research involving human tissue. Cells were grown in Eagle's minimal essential medium (MEM; Invitrogen) with 10% fetal bovine serum (Thermo Scientific) and with 1? penicillin/streptomycin (Thermo Scientific) at 37? C. in a humidified environment containing 5% CO2. Human donor RPE cells were seeded on collagen-coated plates. On day 6 after plating, cells were washed twice with serum-free, phenol red-free MEM (SF-MEM), and treated with HQ in the presence or absence of peptide drugs in SF-MEM for various times at 37? C.

[0081] For WST assay, RPE cells in triplicate wells of a 96-well plate were treated with differing concentrations of HQ (ranging from 140 ?M to 170 ?M) for 2.5 hours in the presence either: No Treatment (control); 400 ?M Risuteganib (RSG) (Positive Control) or Test Compounds (ALG-3001 through ALG-3009) at concentrations of 100 ?M, 400 ?M and 800 ?M. One of the Test Compounds, ALG-3002, was ALG-3002 was additionally tested at concentrations of 12.5 ?M, 25 ?M, 50 ?M and 100 ?M.

[0082] Following the initial 2.5. hour incubation, the medium was removed and the cells were incubated with WST-1 solution for 30 minutes at 37? C. A colorimetric assay was performed based on the cleavage of the tetrazolium salt WST-1 by mitochondrial dehydrogenases in viable cells. The plate was read on a spectrophotometer at 440 nm with a reference wavelength at 690 nm.

[0083] For ROS assay, RPE cells in triplicate wells of 96-well black plates with clear bottoms were washed with SF-MEM, loaded with 20 ?M CM-H2DCFDA in SF-MEM for 30 minutes at 37? C. and then washed twice. Cells were then treated with HQ (160 ?M) in the presence or absence of peptide drugs. Fluorescence was measured at various times with a fluorescence plate reader (490 nm excitation, 522 nm emission).

[0084] For mitochondria membrane potential measurement, RPE cells in triplicate wells of 96-well black plates with clear bottoms were washed with SF-MEM, loaded with 10 ?M JC-1 dye in SF-MEM for 30 minutes at 37? C. and then washed twice. Cells were then treated with HQ (160 ?M) with or without RSG (0.4 mM). A fluorescence plate reader was used to measure the fluorescence at various times to quantify green JC-1 monomer (490 nm excitation, 522 nm emission) and red JC-1 aggregates (535 nm excitation, 590 nm emission).

[0085] For glutathione (GSH) assay, RPE cells in triple wells of 96-well plate were washed with SF-MEM, treated with HQ with and without peptide drugs. After 1.5 hours treatment, cell medium was removed and GSH-Glo was added for 30 minutes incubation at room temperature. Next, detection reagent was added for 15 minutes incubation at room temperature. A plate reader was used to measure the relative luminescence unit, followed by LDH assay through collection of supernatant.

[0086] Data are expressed as the mean?standard deviation. Two-way ANOVA with Tukey multiple comparisons correction was used to determine whether there were statistically significant differences between treatment groups. Results were plotted using GraphPad Prism 9.0 with asterisks indicating the magnitude of P value (N.S.=not significant, * P?0.05, * P?0.01, *** P?0.001, and **** P?0.0001).

[0087] Results: Results showing the cytoprotective effects of the Test Compounds are summarized in Tables 5 and 6 (below):

TABLE-US-00006 TABLE 5 Increased Cell Viability Observed When RSG (Positive Control) and Test Compounds ALG-3001 (P1) through 3009 (P9) Combined With 140 ?M-170 ?M of HQ Toxin Control VS HQ HQ VS-RSG HQ VS ALG-300X HQ VS ALG-300X HQ 150 ?M & ALG-3001-CELL VIABILITY (% of MEM) Control HQ HQ + 400 ?M RSG HQ + 400 ?M ALG-3001 HQ + 800 ?M ALG-3001 100% 12%**** 24%** 27%*** 37%**** HQ 170 ?M & ALG-3001-CELL VIABILITY 100% 13%**** 29%** 27%**** 53%**** HQ 140 ?M & ALG-3002-CELL VIABILITY (% of MEM) Control HQ HQ + 400 ?M RSG HQ + 25 ?M ALG-3002 HQ + 100 ?M ALG-3002 100% 20%**** 44%**** 123%**** 116%**** HQ 160 ?M & ALG-3002-CELL VIABILITY Control HQ HQ + 400 ?M RSG HQ + 50 ?M ALG-3002 HQ + 100 ?M ALG-3002 100% 11%**** 31%**** 161%**** 163%**** HQ 140 ?M & ALG-3003-CELL VIABILITY (% of MEM) Control HQ HQ + 400 ?M RSG HQ + 400 ?M ALG-3003 HQ + 800 ?M ALG-3003 100% 20%**** 44%**** 50%**** 73%**** HQ 160 ?M & ALG-3003-CELL VIABILITY 100% 11%**** 31%**** 50%**** 54%**** HQ 140 ?M & ALG-3004-CELL VIABILITY (% of MEM) Control HQ HQ + 400 ?M RSG HQ + 400 ?M ALG-3004 HQ + 800 ?M ALG-3004 100% 29%**** 67%*** 94%**** 99%**** HQ 160 ?M & ALG-3004-CELL VIABILITY 100% 20%**** 39%*** 60%**** 90%**** HQ 140 ?M & ALG-3005-CELL VIABILITY (% of MEM) Control HQ HQ + 400 ?M RSG HQ + 400 ?M ALG-3005 HQ + 800 ?M ALG-3005 100% 29%*** 67%*** 82%**** 95%**** HQ 160 ?M & ALG-3005-CELL VIABILITY 100% 20%**** 39%*** 36%*** 70%**** HQ 140 ?M & ALG-3006-CELL VIABILITY (% of MEM) Control HQ HQ + 400 ?M RSG HQ + 400 ?M ALG-3006 HQ + 800 ?M ALG-3006 100% 33%**** 61%**** 83%**** 113%**** HQ 160 ?M & ALG-3006-CELL VIABILITY 100% 14%**** 31%** 39%***8 66%**** HQ 140 ?M & ALG-3007-CELL VIABILITY (% of MEM) Control HQ HQ + 400 ?M RSG HQ + 400 ?M ALG-3007 HQ + 800 ?M ALG-3007 100% 33%**** 61%*** 81%**** 97%**** HQ 160 ?M & ALG-3007-CELL VIABILITY 100% 14%**** 31%** 45%**** 73%**** HQ 140 ?M & ALG-3008-CELL VIABILITY (% of MEM) Control HQ HQ + 400 ?M RSG HQ + 400 ?M ALG-3008 HQ + 800 ?M ALG-3008 100% 41%**** 75%*** 77%*** 81%**** HQ 160 ?M & ALG-3008-CELL VIABILITY 100% 26%**** 40%* 38% NS 65%**** HQ 140 ?M & ALG-3009-CELL VIABILITY (% of MEM) Control HQ HQ + 400 ?M RSG HQ + 400 ?M ALG-3009 HQ + 800 ?M ALG-3009 100% 41%**** 75%**** 74%*** 69%*** HQ 160 ?M & ALG-3009-CELL VIABILITY 100% 26%**** 40%**** 52%**** 74%**** In the above experiments HQ + 40 ?M of RSG increases the Cell Viability of the cells above the levels dimished by HQ exposure. In the above experiments HQ + 400 ?M of ALG-3001 to ALG-3009, and HQ + 800 ?M of ALG-3001 to ALG-3009 increase significantly the Cell Viability of the cells above the levels dimished by HQ exposure. (N.S. = not significant, *P ? 0.05, **P ? 0.01, ***P ? 0.001, and ****P ?0.0001

[0088] Table 6 (below) compares the amino acid sequence and highest observed RPE cell viability in HQ-stressed cells for each Test Compound (ALLEG-3001-3009 (P1-P9) as well as the positive control risuteganib (RSG).

TABLE-US-00007 TABLE6 HighestRPE Compound AminoAcidSequence AminoAcidCode Viability ALG- Gly-Arg-Ala-Cys(acid)- GRACys(acid)TP 53% 3001(P1) Thr-Pro ALG- Gly-Arg-Gly-Cys-Thr-Pro GRGCTP 163% 3002(P2) ALG- Gly-Arg-Asp-Gly-Thr-Pro GRDGTP 73% 3003(P3) ALG- Gly-Arg-Ala-Glu-Thr-Pro GRAETP 99% 3004(P4) ALG- Gly-Arg-Gly-Asn-Thr-Pro GRGNTP 95% 3005(P5) ALG- Gly-Arg-Gly-Cys(acid)- GRGCys(acid)GGGDG 113% 3006(P6) Gly-Gly-Gly-Asp-Gly ALG- Gly-Asp-Gly-Arg-Thr-Pro GDGRTP 97% 3007(P7) ALG- Gly-Cys(acid)-Gly-Arg- GCys(acid)GRTP 81% 3008(P8) Thr-Pro ALG- Gly-Lys-Gly-Asp-Thr-Pro GKGDTP 74% 3009(P9) Risuteganib Gly-Arg-Gly-Cys(acid)- GRGCys(acid)TP N/A Thr-Pro

[0089] These results are also summarized graphically in the following figures: FIGS. 1C and 1DALG-3001 (P1); FIGS. 2A, 2B, 2C and 2DALG-3002 (P2); FIGS. 3A and 3BALG-3003 (P3)); FIGS. 4A and 4BALG-3004 (P4); FIGS. 5A and 5B(ALG-3005 (P5); FIGS. 6A and 6B(ALG-3006 (P6); FIGS. 7A and 7BALG-3007 (P7); FIGS. 8A and 8B(ALG-3008 (P8); and FIGS. 9A and 9BALG-3009 (P9).

[0090] It was additionally determined that ALG-3002 (P2) was not only protective against HQ-induced stress but also against CCCP-induced stress. Table 7 (below) and FIG. 10 compare the effects of 100 uM ALG 3002 (P2) in RPE cells stressed with either CCCP or HQ. ALG-3002 (P2) was cytoprotective in both CCCP-stressed cells and HQ-stressed cells.

TABLE-US-00008 TABLE 7 ALG-3002 RPE Cell Viability Against HQ and CCCP Stress in RPE Cells P2 VS Control HQ VS Control HQ VS P2 CCCP VS Control CCCP VS P2 Control ALG-3002 HQ HQ + ALG-3002 CCCP CCCP + ALG- (P2) (P2) 3002 91% 94% NS 38%**** 103%**** 41%**** 88%**** In cells exposed to HQ or CCCP alone Cell Viability is 41%. HQ + 400 ?M ALG-3002 (P2) Cell Viability is 108%. CCCP + 400 ?M of ALG-3002 (P2) Cell Viability is 88%. (N.S. = not significant, *P ? 0.05, **P ? 0.01, ***P ? 0.001, and ****P ? 0.0001

[0091] RPE cell degeneration is central to the pathogenesis of age-related macular degeneration (AMD), a disease that leads to progressive loss of visual function and blindness. Risuteganib, along with these nine new peptides showed an in vitro signal indicating they can preserve RPE cell viability, reduce ROS level and improve mitochondria integrity in human RPE cells stressed by an oxidant associated with macular degeneration. These peptides could be beneficial in the treatment of AMD and other degenerative eye diseases.

[0092] All nine of the Test Compounds protected cell viability, with ALG-3004 (P4), ALG-3006 (P6) and ALG-3007 (P7) providing a greater cytoprotective effect than RSG (positive control). Of the nine Test Compounds, ALG-3002 (P2) showed the greatest cytoprotective effect in this study, which appears to be dose related.

Reactive Oxygen Species (ROS)

[0093] ROS level is a measure of oxidative stress experienced by cells. CM-H2DCFDA-based ROS assay was used to measure the level of ROS after treatments. ALG-3002 (P2), ALG-3004 (P4) and ALG-3007 (P7) were tested and all three were found to significantly reduce ROS level in retinal pigment epithelium (RPE) co-treated with carbonyl cyanide phenylhydrazone (CCCP) or -stressed retinal pigment epithelium (RPE) cells HQ, as plotted, and summarized in Table 8 (below) and shown graphically in FIG. 11 (ALG-3002 (P2)), FIG. 12 (ALG-3004(P4)) and FIG. 13 (ALG-3007(P7)).

TABLE-US-00009 TABLE 8 Reduction of Reactive Oxygen Species in HQ Stress RPE Cells Exposed to ALG-3002, ALG-3004, and ALG-3007 % Reactive Species in HQ Stressed RPE cells Treated with ALG-3002 HQ VS Control Control VS P2 HQ VS HQ + P2 Control HQ-140 ?M ALG-3002(P2)-100 ?M HQ-140 ?M + ALG-3002(P2)-100 ?M 100% 230%**** 73%* 73%**** % Reactive Species in HQ Stressed RPE cells Treated with ALG-3004 HQ VS Control Control VS P4 HQ VS HQ + P4 Control HQ-140 ?M ALG-3004(P4)-400 ?M HQ-140 ?M + ALG-30049P4)-400 ?M 100% 271%**** 100% NS 158%**** % Reactive Species in HQ Stressed RPE cells Treated with ALG-3007 HQ VS Control Control VS P7 HQ VS HQ + P7 Control HQ-140 ?M ALG-3007(7)-400 ?M HQ-140 ?M + ALG-3007(P7)-400 ?M 100% 282%**** 82% NS 175%** (N.S. = not significant, *P ? 0.05, **P ? 0.01, ***P ? 0.001, and ****P ? 0.0001 Reactive oxygen species were increased by HQ to 230% while HQ + ALG-3002 decreased the reactive oxygen species to 73%. Reactive oxygen species were increased by HQ to 47% while HQ + ALG-3004 decreased the reactive oxygen species to 62%. Reactive oxygen species were increased by HQ to 29% while HQ + ALG-3007 decreased the reactive oxygen species to 79%.

Mitochondrial Membrane Potential

[0094] Mitochondria membrane potential is an indicator of mitochondria integrity, where lower potential signifies deteriorated mitochondria and reduced cell health. ALG-3002, ALG-3004 and ALG-3007 were tested and all three were found to significantly improve mitochondria membrane potential in cells co-treated with HQ, as plotted and also summarized in Table 9 (below) and shown graphically in FIG. 14 (ALG-3002 (P2)), FIG. 15 (ALG-3004(P4)) and FIG. 16 (ALG-3007(P7)).

TABLE-US-00010 TABLE 9 Increase in Mitochondrial Membrane Potential of HQ stressed cells Exposed to ALG-3002 (P2), ALG-3004 (P4), and ALG-3007 (P7) Mitochondrial Membrane potential was reduced by HQ to 25% while HQ + ALG-3002 increased the mitochondrial membrane potential to 91%. % Mitochondrial Membrane Potential by ALG-3002 on HQ Stressed RPE Cells Compared to Control Control VS HQ Control VS (P2) HQ VS (P2) Control HQ-140 ?M ALG-3002 (P2)-100 ?M HQ-140 ?M + ALG-3002(P2)-100 ?M 100% 25%**** 100% NS 91%**** % in Mitochondrial Membrane Potential by ALG-3004 on HQ Stressed RPE Cells Compared to Control Control VS HQ Control VS (P4) HQ VS (P4) Control HQ-140 ?M ALG-3004 (P4)-400 ?M HQ-140 ?M + ALG-3004(P4)-400 ?M 106% 47%**** 110%* 62%**** % Change in Mitochondrial Membrane Potential by ALG-3007 on HQ Stressed RPE Cells Compared to Control Control VS HQ Control VS (P7) HQ VS (P7) Control HQ-140 ?M ALG-3007(P7)-400 ?M HQ-140 ?M + ALG-3007(P7)-400 ?M 100% 29%**** 115%** 79%**** Mitochondrial Membrane potential was reduced by HQ to 20% while HQ + ALG-3004 increased the mitochondrial membrane potential to 55%. Mitochondrial Membrane potential was reduced by HQ to 16% while HQ + ALG-3007 increased the mitochondrial membrane potential to 75%. (N.S. = not significant, *P ? 0.05, **P ? 0.01, ***P ? 0.001, and ****P ? 0.0001

Levels of Glutathione (GSH)

[0095] Glutathione (GSH) is a natural antioxidant that cells use to maintain redox homeostasis. GSH level was measured using GSH-Glo GSH assay after treatments. Cells stressed with HQ showed reduced GSH level, while both RSG and ALG-3002 significantly improved GSH level, as summarized in Table 10 (below) and shown graphically in FIG. 17.

TABLE-US-00011 TABLE 10 The Untreated Control level of Glutathione (GSH) is 9064 RLU/LDH units. The presence of HQ Toxin reduces the level of Glutathione (GSH) is reduced to 597 RLU/LDH units. The 800 ?M RSG has no effect on the normal level of Glutathione (GSH) 8995 Increase in Glutathione (GSH) Antioxidant in HQ Stressed RPE Cells Control 160 ?m HQ 400 ?M RSG 160 ?M-HQ + 400 ?M ALG-3002 160 ?M HQ + 400 ?M RSG 400 ?M-ALG-3002 8995? 597? 8995? 3284? 8657? 8060? RLU/LDH units. The presence of 120 ?M of HQ + 400 ?M RSG lowers the level of Glutathione (GSH) to 3248 RLU/LDH units. The 100 ?M ALG-3002 has no effect on the normal level of Glutathione (GSH) 8657 RLU/LDH units. The presence of 120 ?M of HQ + 400 ?M ALG-3002 increases the level of Glutathione (GSH) to 8060 RLU/LDH units.

Effects of RSG and ALG-3002 on Mitochondrial Ultrastructure in Human RPE Cells

[0096] This study compared the effect of risuteganib (RSG) and ALG-3002 (P2) on the morphology of mitochondria in primary human RPE cells stressed with the cigarette smoke toxin, hydroquinone (HQ). HQ is known to induce oxidative stress in cells, leading to cytotoxicity.

[0097] Methods: Primary human RPE cells cultured in 6-well plate were treated with HQ (180 ?M) in the presence or absence of RSG (800 ?M) and ALG-3002 (100 ?M) for 4 hours. After treatment, media was removed and cells then washed with warm PBS (2 mL/well). Cells were lifted by trypsin-EDTA (600 ?L/well), followed by addition of PBS to dilute the trypsin (1 mL/well). Cells were pelleted and washed with PBS (1 mL/well), then fixed with 3% PFA and 2% glutaraldehyde in PBS for 18 hours at 4 degrees Celsius. Cell pellets were then washed with 1 mL PBS and imaged under a transmission EM system for mitochondria ultrastructure changes by HQ and peptides.

[0098] Results/Discussion: FIGS. 18A through 18C are representative of micrographs showing that, in unstressed control cells (FIG. 25A) normal mitochondria morphology was observed, while exposure of the cells to HQ (FIG. 25B) caused alterations in mitochondrial morphology including mitochondrial swelling, presence of vacuoles, and loss of cristae. Both the positive control RSG (FIG. 25C) and the Test Compound ALG-3002 (P2) (FIG. 25D) appeared to significantly reduce the severity of HQ-induced morphologic changes in mitochondria.

[0099] Conclusions: In these primary human RPE cell cultures, RSG and ALG-3002 both protected mitochondria of RPE cells against morphologically apparent HQ stress.

Therapeutic Uses of ALG-3001-ALG-3009

[0100] Mitochondria, the intracellular organelles comprising the main respiratory machinery in cells, are crucial for energy production and cell homeostasis. Due to a high level of metabolic demand by photoreceptors, RPE cells are enriched with a large mitochondrial population to meet the high-energy needs. Consequently, RPE mitochondrial dysfunction can lead to tissue damage and has been implicated in the development of AMD. In RPE cells from eyes with AMD, damaged, fragmented, and ruptured mitochondria have been observed. mtDNA mutation levels are also elevated in RPE cells of eyes with AMD.

[0101] Mitochondrial respiration plays an important role in cell survival. We evaluated the role of RSG in the regulation of mitochondrial function Mitochondria are major sources of ROS generation that contribute to oxidative stress-mediated cell death. To examine whether RSG reduces oxidative stress-mediated ROS production, we measured ROS levels. As shown, ROS production was significantly increased in HQ-treated cells when compared to control, while RSG co-treatment significantly decreased HQ-induced ROS production. RSG alone did not significantly change ROS level when compared to control.

[0102] Primary mitochondrial disorders may sometimes cause skin manifestations (e.g., rashes, pigmentation abnormalities, acrocyanosis) and primary skin disorders may sometimes be linked to mitochondrial dysfunction. A number of skin disorders (e.g., pruritis, atopic dermatitis, psoriasis) may benefit from treatment, as described herein, to improve mitochondrial function. Mitochondrial dysfunction has been characterized as the rule rather than the exception in skin diseases. Feichtinger, R.G., et al.; Mitochondrial dysfunction: a neglected component of skin diseases; Experimental Dermatology Vol. 23, Issue 9, Sep. 2014 (607-614); https://doi.org/10.1111/exd.12484

[0103] Based on the data set forth above, in addition to anti-neovascularization effects described in incorporated U.S. patent applications Ser. No. 16/882,656 entitled PEPTIDE COMPOSITIONS AND RELATED METHODS and Ser. No. 16/882,660 entitled PEPTIDE COMPOSITIONS AND RELATED METHODS, the peptide treatments described herein may be used to treat a variety of disorders, including but not necessarily limited to the following: [0104] Ocular: diabetic retinopathy, retinal neovascularization, exudative or wet macular degeneration, nonexudative or dry macular degeneration, retinitis pigmentosa, glaucoma, other retinal degenerations, dry eye disease, retinal inflammation, scleral inflammation, episcleral inflammation or corneal inflammation. [0105] Neurological: Multiple Sclerosis, Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), Amyotrophic lateral sclerosis (ALS), Peripheral neuropathy, Peripheral nerve pain, stroke, tinnitus and other disorders which cause degeneration of function and/or structure in central and/or peripheral neurons; [0106] Pulmonary: Chronic Obstructive Pulmonary Disease, Pulmonary Fibrosis; [0107] Cardiovascular/Renal: Congestive Heart Failure, Chronic heart failure with reduced ejection fraction, Chronic heart failure with preserved ejection fraction, Barth syndrome, Kidney Failure, Kidney Disease, Kidney failure due to percutaneous renal angiography for renal artery stenosis, Vascular inflammations, Vasculitis, Hemorrhoids; [0108] Hepatologic: Non-Alcoholic Steato Hepatitis (NASH) [0109] Muscular: Impaired skeletal muscle function in the elderly, primary muscle mitochondrial myopathy or neuropathy, ischemia-reperfusion injury, Cardiac or skeletal muscle impairment resulting from protozoal or other microbial infections; [0110] Dermatological/Topical: Rash, Pigmentation abnormality, Acrocyanosis, Atopic Dermatitis, Malasma, Pruritis, Psoriasis and Hemorrhoids; [0111] Otic: inflammations of the middle or inner ear, Meniere's disease, sensorineural hearing loss or tinnitus

[0112] The dosage, dosing schedule and/or route of administration may differ depending on the type and severity of disease or disorder being treated. For example, in at least some applications where a compound comprising ALG-3001 through ALG-3009 is administered intravitreally or topically to a subject's eye the dosage of the compound selected from ALG-3001 to ALG-3009 may be in the following range(s) or others as may be clinically appropriate: [0113] a) Intravitreal injection or implantation: 0.01 mg/50 uL-10.0 mg/50 uL, or 1.0 mg/50 uL-2.0 mg/50 uL [0114] b) Topical application to the eye: ?0.01 g/100 mL-40 g/100 mL, or 0.60 g/100 mL [0115] c) Intravenous injection or infusion: 0.01 mg/kg body weight ?50 mg/kg body weight, or 5.0 mg/kg body weight ?6.0 mg/kg body weight [0116] d) Topical applications to skin or mucous membranes: 0.01 g/100 g-40 g/100 g, or 0.40 g/100g-0.50 g/100 g [0117] e) Intradermal, subcutaneous, intramuscular, intraperitoneal or other injection or implantation: 0.01 mg/kg body weight ?50 mg/kg body weight, or 5.0 mg/kg body weight ?6.0 mg/kg body weight [0118] f) Intreathecal, epidural: 0.01 mg/kg body weight ?50 mg/kg body weight, or 2.0 mg/kg body weight ?6.0 mg/kg body weight [0119] g) Ear (e.g., otic, transtympanic, intratympanic, intracochlear) application for inflammations of the middle or inner ear, Meniere's disease, sensorineural hearing loss or tinnitus: 0.01 g/100 g-40 g/100 g, or 0.40 g/100 g-0.50 g/100 g [0120] h) Rectal or transmucosal application (e.g., suppository or topical application for treatment of vascular inflammation, vasculitis, Hemorrhoids or for systemic transmucosal absorption): 0.01 g/100 g-40 g/100 g, or 0.40 g/100 g-0.50 g/100 g

[0121] Although the above description refers to examples or embodiments, various additions, deletions, alterations and modifications may be made to those described examples and embodiments without departing from the intended spirit and scope of the invention(s) disclosed herein. For example, any elements, steps, members, components, compositions, reactants, parts or portions of one embodiment or example may be removed/eliminated and/or incorporated into or used with another embodiment or example, unless otherwise specified or unless doing so would render that embodiment or example unsuitable for its intended use. Also, where the steps of a method or process have been described or listed in a particular order, the order of such steps may be changed unless otherwise specified or unless doing so would render the method or process unsuitable for its intended purpose. Additionally, the elements, steps, members, components, compositions, reactants, parts or portions of any invention or example described herein may optionally exist or be utilized in the absence or substantial absence of any other element, step, member, component, composition, reactant, part or portion unless otherwise noted. All reasonable additions, deletions, modifications and alterations are to be considered equivalents of the described examples and embodiments and are to be included within the scope of the following claims.