Methods to protect cells from damage

Abstract

Described herein are methods for preventing non-injurious cell damage during exposure of a subject to radiation. In some embodiments, the method can include administering to the subject an amount of poloxamer 188 or a pharmaceutical formulation thereof prior to exposure to the insult.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the benefit and priority of the concurrently pending U.S. Provisional Patent Application No. 63 / 050,075, entitled “METHODS OF PROTECTING CELLS FROM INSULTS,” filed on 9 July 2020, the contents of which are incorporated herein by reference in their entirety.

[0002] Statement on federally funded research This invention was made with government support under grant number OIA-1655740, awarded by the National Science Foundation. The U.S. Government reserves certain rights in this invention.

[0003] The subject matter disclosed herein generally relates to methods for protecting cells, such as non-cancerous cells, from radiation and / or other damage. [Background technology]

[0004] Radiotherapy is a well-established cancer treatment modality that has been used for over 50 years. Radiation directly causes DNA damage in cells, such as single-strand breaks (SSBs), double-strand breaks (DSBs), DNA crosslinks, and DNA-protein crosslinks, or indirectly induces DNA damage by reactive oxygen species (ROS) / reactive nitrogen species (RNS), which leads to cellular senescence and apoptosis. Radiation inherently does not distinguish between cancerous and non-cancerous cells. Although methods and techniques that target only cancerous cells have been developed (e.g., sensitizing cancerous cells to require less radiation dose and physically limiting the area of ​​direct exposure), radiotherapy is still considered a "double-edged sword" and can still cause significant damage to non-cancerous cells.

[0005] Other injuries, such as mechanical, chemical (e.g., hydrogen peroxide or other drugs), or biological (e.g., viruses and microorganisms), can result in ROS, RNS, peroxides, superoxides, or other inflammatory damage in surrounding, and potentially distant, cells and tissues that are not directly exposed to the injury. [Overview of the project] [Problems that the invention aims to solve]

[0006] Therefore, there is still a need for compositions, methods, and techniques to protect adjacent cells and tissues that are not directly exposed to damage resulting in the production of ROS, RNS, peroxides, superoxides, or other inflammatory damage in surrounding, and sometimes distant, cells and tissues.

[0007] Any reference or specification of any document in this application does not constitute an endorsement that such document is available as prior art for the present invention. [Means for solving the problem]

[0008] In certain exemplary embodiments, a method for preventing damage to non-injured cells in a subject during and / or after injury exposure is described herein, the method comprising: administering a first amount of poloxamer 188 (P188) or a pharmaceutical formulation thereof to one or more areas of the subject before exposing them to injury; and exposing one or more areas of the subject to injury after the blood concentration of P188 has reached 1 to 5 mg / mL.

[0009] In certain exemplary embodiments, a first amount of P188 is administered intravenously.

[0010] In a particular exemplary embodiment, the method involves administering a first amount of P188 and before exposing one or more areas of the subject to injury, n th This further includes administering a certain amount of P188 or a pharmaceutical preparation thereof to a target, where n is 2 or greater.

[0011] In certain exemplary embodiments, the first amount of P188, n th amount, or all of the amount, is an amount effective to increase the blood concentration of P188 to 1 - 5 mg / mL within 1 - 48 hours.

[0012] In certain exemplary embodiments, the first amount or n th amount of P188 is administered over a period as a continuous infusion.

[0013] In certain exemplary embodiments, one or more of the first amount or n th amount of P188 is administered as a bolus amount.

[0014] In certain exemplary embodiments, only the first amount is administered, and the first amount is effective to increase the blood concentration of P188 to about 1 - about 5 mg / mL within 1 - 48 hours.

[0015] In certain exemplary embodiments, the subject does not receive an amount of P188 for at least 14 hours after receiving the first or n th amount of P188.

[0016] In certain exemplary embodiments, P188 administration is discontinued immediately after exposing one or more regions of the subject to injury.

[0017] In certain exemplary embodiments, exposing one or more regions of the subject to injury is performed about 1 - about 48 hours after administering the first or n th amount of P188.

[0018] In certain exemplary embodiments, the injury is effective to kill one or more cancerous cells within one or more regions of the subject.

[0019] In certain exemplary embodiments, the non - injured cells are non - cancerous cells.

[0020] In certain exemplary embodiments, the non - injured cells are endothelial cells.

[0021] In certain exemplary embodiments, the injury may be a mechanical injury, a chemical injury, a biological injury, an energetic injury, or a combination thereof.

[0022] In certain exemplary embodiments, the injury is caused by ionizing radiation.

[0023] In certain exemplary embodiments, a method for preventing damage to non-injured cells in a subject during and / or after injury exposure is described herein, the method comprising: administering a certain amount of poloxamer 188 (P188) or a pharmaceutical formulation thereof to one or more areas of the subject before exposure of the subject to injury, the amount being in the range of about 10 to 150 mg / kg; and exposing one or more areas of the subject to injury within 0 to 48 hours after administration of the certain amount of P188 or a pharmaceutical formulation thereof.

[0024] In certain exemplary embodiments, a certain amount of P188 or its pharmaceutical formulation is administered intravenously.

[0025] In certain exemplary embodiments, administration of P188 or its pharmaceutical formulation is discontinued immediately before or after exposure of one or more areas of the subject to injury.

[0026] In certain exemplary embodiments, the subject is not administered any amount of P188 or the pharmaceutical formulation for at least 14 hours after being administered a certain amount of P188 or the pharmaceutical formulation, or after discontinuing the administration of P188 or the pharmaceutical formulation.

[0027] In certain exemplary embodiments, the injury may be a mechanical injury, a chemical injury, a biological injury, an energetic injury, or a combination thereof.

[0028] In certain exemplary embodiments, the injury is caused by ionizing radiation.

[0029] In certain exemplary embodiments, a kit is described herein comprising: a certain amount of P188 or a pharmaceutical formulation thereof, which is effective in preventing damage to non-injured cells in a subject when one or more areas of the subject are exposed to injury; and instructions imposed on a tangible medium of expression that instruct the subject to be administered a certain amount of P188 or a pharmaceutical formulation thereof 0 to 48 hours before one or more areas of the subject are exposed to injury.

[0030] In certain exemplary embodiments, the injury may be a mechanical injury, a chemical injury, a biological injury, an energetic injury, or a combination thereof.

[0031] In certain exemplary embodiments, the injury is caused by ionizing radiation.

[0032] In certain exemplary embodiments, the instructions further instruct the subject to discontinue administration of a certain amount of P188 or its pharmaceutical formulation immediately before or after exposure to an injury in one or more areas of the subject.

[0033] In certain exemplary embodiments, a certain amount of P188 or its pharmaceutical formulation is effective in achieving a blood concentration of approximately 1 to approximately 5 mg / mL in a subject within 0 to 48 hours.

[0034] In certain exemplary embodiments, a method for protecting non-injured cells after exposure of a subject to injury is described herein, the method comprising: a) administering a certain amount of P188 or a formulation thereof to a subject before exposure to injury; b) administering a certain amount of P188 or a formulation thereof to a subject immediately after exposure to injury; c) administering a certain amount of P188 or a formulation thereof to a subject during injury; or d) a combination thereof.

[0035] In certain exemplary embodiments, a certain amount is effective in increasing the blood concentration of P188 in a subject to 1–5 mg / mL.

[0036] In certain exemplary embodiments, a certain amount is effective in coating one or more non-injured cells.

[0037] In certain exemplary embodiments, a certain amount is effective in reducing or preventing oxidative damage to one or more non-injured cells.

[0038] In certain exemplary embodiments, a certain amount is effective in reducing or preventing inflammatory damage to one or more non-injured cells.

[0039] In certain exemplary embodiments, administration is performed 0 to 48 hours before exposure to injury.

[0040] In certain exemplary embodiments, administration is performed 1 to 24 hours after exposure to injury.

[0041] In certain exemplary embodiments, the injury may be a mechanical injury, a chemical injury, a biological injury, an energetic injury, or a combination thereof.

[0042] In certain exemplary embodiments, the injury is caused by ionizing radiation.

[0043] In certain exemplary embodiments, a method for preventing injury-induced pneumonia in a subject is described herein, the method being: a) Administering a certain amount of P188 or a preparation thereof to the subject before exposure to injury; b) Administering a certain amount of P188 or a preparation thereof to the subject immediately after exposure to injury; c) Administering a certain amount of P188 or a preparation thereof to the subject during injury; or d) combinations of those Includes.

[0044] In certain exemplary embodiments, a certain amount is effective in increasing the blood concentration of P188 in a subject from 1 mg / mL to 5 mg / mL.

[0045] In certain exemplary embodiments, a certain amount is effective in coating one or more non-injured cells.

[0046] In certain exemplary embodiments, a certain amount is effective in reducing or preventing oxidative damage to one or more non-injured cells.

[0047] In certain exemplary embodiments, a certain amount is effective in reducing or preventing inflammatory damage to one or more non-injured cells.

[0048] In certain exemplary embodiments, administration is performed 0 to 48 hours before exposure to injury.

[0049] In certain exemplary embodiments, administration is performed 0 to 24 hours after exposure to injury.

[0050] In certain exemplary embodiments, the injury may be a mechanical injury, a chemical injury, a biological injury, an energetic injury, or a combination thereof.

[0051] In certain exemplary embodiments, the injury is caused by ionizing radiation.

[0052] In certain exemplary embodiments, a method for treating a disease or disorder in an object requiring treatment is described herein, the method being: a) Administering a certain amount of P188 or a preparation thereof to the subject before exposure to injury; b) Administering a certain amount of P188 or a preparation thereof to the subject immediately after exposure to injury; c) Administering a certain amount of P188 or a preparation thereof to the subject during injury; or d) combinations of those Includes.

[0053] In certain exemplary embodiments, a certain amount is effective in increasing the blood concentration of P188 in a subject from 1 mg / mL to 5 mg / mL.

[0054] In certain exemplary embodiments, a certain amount is effective in coating one or more non-injured cells.

[0055] In certain exemplary embodiments, a certain amount is effective in reducing or preventing oxidative damage to one or more non-injured cells.

[0056] In certain exemplary embodiments, a certain amount is effective in reducing or preventing inflammatory damage to one or more non-injured cells.

[0057] In certain exemplary embodiments, administration is performed 0 to 48 hours before exposure to injury. In certain exemplary embodiments, administration is performed 0 to 24 hours or 0 to 48 hours after exposure to injury. In certain exemplary embodiments, P188 is not administered before and / or during exposure to injury.

[0058] In certain exemplary embodiments, the injury may be a mechanical injury, a chemical injury, a biological injury, an energetic injury, a physiological injury, or a combination thereof.

[0059] In certain exemplary embodiments, the injury is an ischemic event, possibly a stroke or myocardial infarction.

[0060] These and other aspects, purposes, features, and advantages of the exemplary embodiments will become apparent to those skilled in the art by considering the following detailed description of the exemplary embodiments.

[0061] An understanding of the features and advantages of the present invention will be obtained by referring to the following detailed description illustrating exemplary embodiments in which the principles of the present invention may be utilized, and to the accompanying drawings. [Brief explanation of the drawing]

[0062] [Figure 1] This panel shows fluorescence microscopy images from in vitro studies that may demonstrate Hiroloxamer protects endothelial cells from hydrogen peroxide.

[0063] [Figure 2A-2B]The graph shows results from in vitro studies that may demonstrate that Hiroloxamer does not affect the sensitivity of cancer cells to ionizing radiation, as modeled by the A549 (Figure 2A) and H460 (Figure 2B) human lung cancer cell lines.

[0064] [Figure 3A-3P] The images show fluorescence microscopy from an in vitro study demonstrating that Hiroloxamer preserves the actin cytoskeleton of human adipose microvascular endothelial cells directly exposed to ionizing radiation.

[0065] [Figure 4A-4E] Bright-field images (Figures 4A-4D) and graphs (Figure 4E) from an in vitro study are shown, illustrating the cell viability within the irradiated area (n=16).

[0066] [Figures 5A-5E] Bright-field images (Figures 5A-5D) and graphs (Figure 5E) from an in vitro study are shown, illustrating the survival rate in areas adjacent to the irradiated region (n=9).

[0067] [Figures 6A-6E] Bright-field images (Figures 6A-6D) and graphs (Figure 6E) from an in vitro study are shown, illustrating the survival rate in the region furthest from the irradiated area (n=4).

[0068] [Figure 7A-7L] Fluorescence images from an in vitro study showing pre-angiogenic endothelial fibroblast constructs without scaffolds, exposed to 10 Gy of X-ray irradiation, are shown. Figures 7A-7D show control, no Rx, and no radiation (73.655 μm with a 3.51 μm step size). Figures 7E-7H show exposure to 10 Gy and no Rx (76.027 μm with a 4 μm step size). Figures 7I-7L show Rx pretreatment and exposure to 10 Gy (88.031 μm with a 4 μm step size). Hoescht (nucleus), Phalloidin (F-actin), and CD31 (endothelial cell network) are shown in grayscale.

[0069] [Figures 8A-8C] This specification describes several embodiments of methods for protecting cells, particularly non-cancerous cells, from radiation using the P188 formulation described herein.

[0070] [Figures 9A-9C] Fluorescence images are shown from in vitro studies that may demonstrate the effects of Hiorloxamer pretreatment on F-actin and the endothelial network. SPEC control: The EC network is a primitive capillary-like network. Control SPEC(6): 73.655 μm with a 3.51 μm step size (22 steps). No Rx, XRT SPEC(3): 76.027 μm with a 4 μm step size (28 steps). Rx pretreatment, full exposure SPEC(1): 88.031 μm with a 4 μm step size (23 steps).

[0071] [Figure 10A-10D] The following fluorescence images show results from an in vitro study that may demonstrate the effect of pretreatment with Hiroloxamer on cells exposed to 1 Gy of radiation.

[0072] [Figure 11] Fluorescence images are shown showing results from an in vitro study, which may indicate that 2Gy exposure can lead to intracellular gap formation, and that this can be mitigated by pretreatment with Hiroloxamer. Tension fibers are required to induce cell contraction and dramatically influence the rate and size of interendothelial gaps formed when cells retract from their boundaries (Pasain). Direct association of actin cytoskeleton with cell adhesion proteins is essential for barrier function. Tight junctions and adherent junctions connect adjacent cells and regulate paracellular permeability. Without Rx: Intercellular gap formation indicates a loss of cell-cell junctions, leading to F-actin depolymerization in Ecs.

[0073] [Figure 12]The image shows fluorescence results from an in vitro study that may demonstrate the effect of pretreatment with Hiroloxamer on cells exposed to 50 Gy of radiation.

[0074] [Figure 13] This document presents lung histology results from an in vivo study of PBS- or hiroloxamer-treated rats, demonstrating the potential effects of 200 mg / kg hiroloxamer treatment three hours prior to 20 Gy irradiation. Lung samples for histology were collected approximately 24 hours after irradiation.

[0075] [Figure 14] This shows lung histology results from an in vivo study of PBS-treated or hiroloxamer-treated rats, demonstrating the effect of 200 mg / kg hiroloxamer treatment administered 15 minutes prior to exposure to 20 Gy of irradiation.

[0076] [Figure 15] Lung histology is shown from an in vivo study of PBS or hiroloxamer-treated rats, demonstrating the effect of 200 mg / kg hiroloxamer treatment administered 15 minutes prior to exposure to 20 Gy irradiation. Histological references of healthy controls were obtained from http: / / histology.oucreate.com / Captions / Respiratory / 109.ling.mammal / 109.bronchiole.c1.40.Lhtm#click.

[0077] [Figure 16] This shows lung histology results from an in vivo study of PBS-treated or hiroloxamer-treated rats, demonstrating the effect of 200 mg / kg hiroloxamer treatment administered 15 minutes prior to exposure to 20 Gy of irradiation.

[0078] [Figure 17]This shows lung histology results from an in vivo study of PBS-treated or hiroloxamer-treated rats, demonstrating the effect of 200 mg / kg hiroloxamer treatment administered 15 minutes prior to exposure to 20 Gy of irradiation.

[0079] [Figure 18] This image shows lung histology from an in vivo study of PBS-treated rats, demonstrating that oxygen pathways can be preserved in the bronchioles in lungs irradiated with Hiroloxamer. A histological reference of healthy controls was obtained from http: / / histology.oucreate.com / Captions / Respiratory / 109.ling.mammal / 109.bronchiole.c1.40.Lhtm#click.

[0080] [Figure 19] This shows lung histology results from an in vivo study of PBS-treated or hiroloxamer-treated rats, demonstrating the effect of 200 mg / kg hiroloxamer treatment administered 15 minutes prior to exposure to 20 Gy of irradiation.

[0081] [Figure 20] We present lung histology results from an in vivo study of PBS-treated or Hiroloxamer-treated rats, which may demonstrate that Hiroloxamer reduces bronchiolar inflammation through vascular protection.

[0082] [Figure 21] This paper presents lung histology results from in vivo studies of PBS-treated and Hiroloxamer-treated rats, as well as healthy controls, demonstrating that Hiroloxamer can preserve vascular (V) and bronchiolar (B) structures.

[0083] [Figure 22]This figure shows lung histology results from in vivo studies of PBS and Hiroloxamer-treated rats, demonstrating that Hiroloxamer can preserve vascular (V) and bronchiolar (B) structures. Figure 22 is a high-magnification image of the same as shown in Figure 21.

[0084] [Figure 23] Lung histology is shown from in vivo studies of PBS and Hiroloxamer-treated rats, demonstrating that Hiroloxamer can preserve vascular (V) and bronchiolar (B) structures. Healthy controls are also shown.

[0085] [Figure 24] This paper presents lung histology results from in vivo studies of PBS and Hirorloxamer, suggesting that Hiroloxamer may be able to attenuate early radiation-induced injury in the lungs.

[0086] [Figure 25] This paper presents lung histology results from an in vivo study of PBS-treated and Hiroloxamer-treated rats, demonstrating that Hiroloxamer can preserve the perivascular space of the irradiated lungs.

[0087] [Figure 26] This paper presents lung histology results from an in vivo study of PBS-treated and Hiroloxamer-treated rats, demonstrating that Hiroloxamer can preserve the perivascular space of the irradiated lungs.

[0088] [Figure 27] The image shows lung histology from an in vivo study of PBS and Hiroloxamer-treated rats, demonstrating that Hiroloxamer can preserve the perivascular tissue of the irradiated lung. Figure 27 is a higher magnification of the image shown in Figure 26.

[0089] [Figure 28] The irradiation protocols for the results shown in Figures 13-27 are presented.

[0090] [Figure 29] Figures 13-27 show an overview of the animal populations used in these experiments.

[0091] [Figure 30] The results from an in vivo study one week after irradiation (20 Gy) are shown. Figure 30 shows a panel of representative micrographs of hematoxylin and eosin (H&E) stained lung sections one week after irradiation from the control, radiation (RT), and Hiroloxamer+ radiation groups (scale bar = 250 μm). The control group had normal (clear) lumens of the lung bronchi ("Br") and normal lumens of the air alveoli. The RT group clearly had inflammatory infiltration to almost obstruction throughout the entire peribronchial region, as well as collapsed blood vessels and capillaries, thickening of the alveolar septa, and cellular infiltration surrounding the bronchial structures. The Hiroloxamer+RT group had mild inflammation around the major perivascular vessels of the peribronchial region, but the alveoli and perivascular regions appeared normal.

[0092] [Figure 31] Results from an in vivo study 6 weeks post-irradiation (20 Gy) suggest that Hioloxamer may be able to preserve healthy lung tissue in a partial-volume rat lung X-ray irradiation model. Figure 31 shows representative micrographs of hematoxylin and eosin (H&E) stained right lower lung lobe sections from control, radiotherapy (RT), and Hiroloxamer + radiotherapy 6 weeks post-irradiation (scale bar = 250 μm). The control group had normal (clear) lumens of the lung bronchi ("Br") and normal lumens of the air alveoli. The RT group had marked interstitial edema, vascular and capillary ("V") congestion, increased thickness of the alveolar septa, and dense inflammatory infiltration throughout the entire lobe distally dilated in the right upper lung lobe. The Hiroloxamer + RT group had several inflammatory infiltrations surrounding several bronchioles located near some minor alveolar thickenings, but the blood vessels remained intact and the surrounding inflammatory infiltration was minimal.

[0093] [Figure 32]This image shows a representative micrograph of a picrosilius red-stained right lower lung lobe section from an in vivo study 6 weeks after irradiation with 20 Gy of X-rays (scale bar = 250 microns).

[0094] [Figure 33] Representative micrographs of hematoxylin and eosin (H&E) stained right upper lung lobe sections from control, radiation (RT), and Hiroloxamer+ radiation studies 6 weeks post-irradiation are shown (scale bar = 250 microns). Radiation-induced lung injury spread from the radiation-exposed right lower lung lobe to the distal right upper lung lobe only in the 20 Gy RT group.

[0095] [Figure 34] Representative images and micrographs are shown to demonstrate that Hiroloxamer may prevent the development of acute pneumonia in a rat model of radiation-induced pneumonia. Exposure of SD rats to a single dose of 20 Gy of X-ray irradiation is the prescription dose for radiation-induced pneumonia (Ghita). The rats were anesthetized and restrained in a custom-made jig designed to expose only 6 mm of the right central lung lobe to irradiation, while the rest of the animals remained shielded by a lead shield. To examine the development and progression of pneumonia, proximal and distal tissues of the irradiated site were stained with hematoxylin and eosin (H&E), and the integrity of major lung structures was assessed 6 weeks post-irradiation. Representative micrographs of right lower lung lobe sections from healthy normal, vehicle (saline) + RT, and Hiroloxamer + RT rats 6 weeks post-irradiation. Vehicle (saline) + RT control animals developed acute pneumonia within 6 weeks post-irradiation, as evidenced by capillary leakage, marked interstitial edema, vascular and capillary congestion, increased alveolar thickness, and dense inflammatory infiltration distally propagating to adjacent lobes and the contralateral left lung. Animals treated with Hiroloxamer before irradiation (Hiroloxamer + RT) had clear peribronchial and bronchial lumens and appeared normal alveolar septa, similar to healthy control animals (healthy normal); n=3 for each group.

[0096] [Figures 35A-35D]Representative micrographs (Figures 35A-35C) and graphs (Figure 35D) illustrating MPO analysis of leukocyte infiltration are shown. Controlled neutrophil degranulation and myeloperoxidase (MPO) release at the injury site are necessary for effective wound healing. However, excessive degranulation can exaggerate the inflammatory response and lead to tissue damage even in the absence of infection. The level of MPO activity indicates the state of inflammation and oxidative stress in the tissue. To model radiation-induced pneumonia, SD rats were irradiated with a single 20 Gy dose of X-rays through a 6 mm lead shield in the right central lung lobe, as previously described. Sections from three right and contralateral left lung lobes were paraffin-embedded, sectioned to 5 μm, and stained with the Hanker-Yates peroxidase leukocyte kit (Sigma Aldrich) for analysis. Stained sections were imaged using a LionHeartFX automated microscope, and MPO+ cell counting was automated via thresholding in Gen5 software. Three animals were used in each group, and five high-power fields were analyzed for each animal. (Figure 35A) Representative MPO-stained right lower lung lobe sections of healthy controls, (Figure 35B) vehicle (saline) + RT, and (Figure 35C) 200 mg / kg Hiroloxamer + RT. Tissue damage was observed in the control saline + RT animals, and they had significantly elevated MPO levels in all lung lobes compared to Hiroloxamer + RT and healthy normal animals (Figure 35D). Statistical comparisons (ANOVA, followed by Tukey) of healthy normal animals, vehicle (saline) + RT, and Hiroloxamer + RT showed no difference between healthy normal animals and Hiroloxamer + RT, while there were significant statistical differences between vehicle (saline) + RT and healthy normal animals (p<0.001), and between vehicle (saline) + RT and Hiroloxamer + RT (p<0.001).

[0097] [Figures 36A-36B]This graph shows that radiotherapy, in combination with dose-escalating hiroloxamer, effectively kills human cancer cells. A549 and H460 human cancer cell lines were treated with hiroloxamer before single-dose irradiation of 0, 2, or 6 Gy. No significant survival trend was observed with dose-escalating hiroloxamer treatment.

[0098] [Figure 37] Representative microscopic images are shown that demonstrate Hiroloxamer protects healthy tissue from radiotoxicity regardless of radiation dose. Animals were irradiated with either 10 Gy × 1 fx, 20 Gy × 1 fx, or 10 Gy × 4 fx X-rays at 2-3 days intervals for 2 weeks in an 8 mm area of ​​the right lung. All animals were sacrificed 6 weeks after irradiation and stained with H&E (5 μm sections). Animals treated with the vehicle (upper panel of the figure) had varying degrees of apparent radiation-induced lung injury based on the dose. The 20 Gy × 1 fx dose resulted in the most significant injury, which was evidenced by edema, inflammatory infiltration, and congestion of the lung parenchyma, which were not localized to the irradiation but spread to all lobes on both sides of the lung. These histological findings indicate the development of radiation-induced acute pneumonia. 10 Gy × 1 fx and 10 Gy × 4 fx caused less damage than 20 Gy × 1 fx, which is consistent with the classical theory that healthy tissue toxicity depends on the dose per dose rather than the total dose delivered. Animals pre-treated with 200 mg / kg of Hiroloxamer showed only localized damage in the targeted irradiation area, and healthy tissue was protected from toxicity in all radiotherapy regimens evaluated.

[0099] [Figures 38A-38D]Representative microscopic images are shown that may demonstrate dose-dependent protection of healthy tissue from radiotoxicity by Hiroloxamer. Animals were exposed to 10 Gy x 4 doses of X-ray irradiation delivered every 2-3 days over 2 weeks in an 8 mm area of ​​the right lung. Animals treated with 200 mg / kg of Hiroloxamer before irradiation (Figure 38D) had a normal lung appearance comparable to that of healthy controls (Figure 38A). Animals treated with 50 mg / kg of Hiroloxamer (Figure 38C) had evident cellular infiltration and subsequent alveolar thickening, which was not as severe as in vehicles (PBS) treated animals (Figure 38B), but did not provide adequate protection compared to the 200 mg / kg dose. [Modes for carrying out the invention]

[0100] The drawings in this specification are for illustrative purposes only and are not necessarily drawn to scale.

[0101] Before describing this disclosure in more detail, it should be understood that this disclosure is not limited to the specific embodiments described and is therefore naturally subject to change. It should also be understood that the terms used herein are intended solely to describe specific embodiments and are not intended to limit them.

[0102] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this disclosure belongs. Any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of this disclosure, but preferred methods and materials are described herein.

[0103] All publications and patents cited herein are cited to disclose and describe the relevant methods and / or materials from which the publications are cited. All such publications and patents are incorporated herein by reference as if it were specifically and individually indicated that each individual publication or patent is incorporated by reference. Such incorporation by reference is expressly limited to the methods and / or materials described in the cited publications and patents, and does not extend to the lexicographical definitions from the cited publications and patents. Furthermore, lexicographical definitions in the cited publications and patents that are not expressly repeated in this application should not be treated as such and should not be read as defining any term appearing in the attached claims. Any citation of a publication relates to its disclosure prior to the filing date, and should not be construed as an acknowledgment that this disclosure does not grant prior rights to such publications by prior disclosure. Furthermore, the dates of the publications provided may differ from the actual publication dates that may need to be independently verified.

[0104] As will be apparent to those skilled in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has distinct components and features that can be readily separated from or combined with features of any of several other embodiments without departing from the scope or spirit of this disclosure. Any of the listed methods may be performed in the order of the listed events, or in any other logically possible order.

[0105] Where a range is expressed, further embodiments include a range from one specific value to and / or another specific value. Enumeration of numerical ranges by endpoints includes all numbers and fractions contained within each range, as well as the enumerated endpoints. Where a range of values ​​is provided, unless the context specifically indicates otherwise, each intervening value up to one-tenth of the lower limit between the upper and lower limits of that range, and any other stated or intervening values ​​within that stated range, are understood to be included in this disclosure. The upper and lower limits of these smaller ranges may independently be contained within smaller ranges and are also included in this disclosure, subject to any specifically excluded limits within the stated range. Where a stated range includes one or both limits, ranges excluding either or both of those included limits are also included in this disclosure. For example, if a described range includes one or both of the limits, the disclosure also includes ranges that exclude either or both of the limits that they include. For example, the phrase "x~y" includes the range from "x" to "y" and the range greater than "x" and less than "y". A range can also be expressed as an upper limit, for example, "about x, y, z, or less," which should be interpreted as including the specific ranges of "about x," "about y," and "about z," as well as the ranges of "less than x," "less than y," and "less than z." Similarly, the phrase "about x, y, z, or greater than" should be interpreted as including the specific ranges of "about x," "about y," and "about z," as well as the ranges of "greater than x," "greater than y," and "greater than z." In addition, the phrase "about 'x'~'y'" includes "about 'x'~about 'y'" when 'x' and 'y' are numerical values.

[0106] It should be noted that ratios, concentrations, quantities, and other numerical data may be expressed in range form as specified herein. It will be further understood that each endpoint of a range has significant meaning in relation to and independently of the other endpoint. It will also be understood that numerous values ​​are disclosed herein, and each value is disclosed herein not only as the value itself but also "about" that particular value. For example, if the value "10" is disclosed, "about 10" is also disclosed. Ranges may be expressed herein as "about" one particular value and / or "about" another particular value. Similarly, it will be understood that when a value is expressed as an approximation by the use of the antecedent "about," the particular value forms further aspects. For example, if the value "about 10" is disclosed, "10" is also disclosed.

[0107] Such range formats are used for convenience and brevity, and should therefore be interpreted flexibly to include not only the numbers explicitly listed as the limits of the range, but also all individual numbers or subranges contained within that range, as if each number and subrange were explicitly listed. For example, the numerical range "approximately 0.1% to 5%" should be interpreted to include not only the explicitly listed values ​​of approximately 0.1% to approximately 5%, but also the individual values ​​within the indicated range (e.g., approximately 1%, approximately 2%, approximately 3%, and approximately 4%) and subranges (e.g., approximately 0.5% to approximately 1.1%; approximately 5% to approximately 2.4%; approximately 0.5% to approximately 3.2%, and approximately 0.5% to approximately 4.4%, as well as other possible subranges).

[0108] general definition Unless otherwise defined, technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which this disclosure pertains. For definitions of general terms and techniques in molecular biology, see Molecular Cloning: A Laboratory Manual, 2 ndedition(1989)(Sambrook,Fritsch,and Maniatis);Molecular Cloning:A Laboratory Manual,4 th edition(2012)(Green and Sambrook);Current Protocols in Molecular Biology(1987)(F.M.Ausubel et al.eds.);the series Methods in Enzymology(Academic Press,Inc.):PCR 2:A Practical Approach(1995)(M.J.MacPherson,B.D.Hames,and G.R.Taylor eds.):Antibodies,A Laboratory Manual(1988)(Harlow and Lane,eds.):Antibodies A Laboratory Manual,2 ndedition 2013(EAGreenfield ed.);Animal Cell Culture(1987)(RIFreshney,ed.);Benjamin Lewin,Genes IX,published by Jones and Bartlet,2008(ISBN 0763752223);Kendrew et al.(eds.),The Encyclopedia of Molecular Biology,published by Blackwell Science Ltd.,1994(ISBN 0632021829); Robert A. Meyers(ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by VCH Publishers, Inc., 1995(ISBN 9780471185710); Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons(New York, NY1994), March, Advanced Organic Chemistry Reactions, Mechanisms and Structure 4th ed., John Wiley & Sons (New York, NY1992); and Marten H. Hofker and Jan van Deursen, Transgenic Mouse Methods and Protocols, 2 nd It can be found in the 2011 edition.

[0109] As used herein, the singular forms “a,” “an,” and “the” refer to both singular and plural objects unless the context clearly indicates otherwise.

[0110] As used herein, “about,” “approximately,” “substantially,” etc., when used in relation to measurable variables such as parameters, quantities, durations, etc., mean to include variations of a particular value and from that value, e.g., variations of + / -10%, + / -5%, + / -1%, and + / -0.1% or less of the particular value and from that value, including experimental errors (e.g., which can be determined by a given dataset, a technically acceptable standard, and / or e.g., a given confidence interval (e.g., a 90%, 95%, or higher confidence interval from the mean)) (to the extent that such variations are appropriate for the work to be performed in the disclosed invention). As used herein, the terms “about,” “approximately,” “to, or about,” and “substantially” may mean that the quantity or value in question may be an exact value or a value that provides an equivalent result or effect as described in the claims or taught herein. That is, quantities, sizes, formulations, parameters, and other quantities and characteristics are not, and do not need to be, exact, and may be approximate and / or greater or less, as desired, to reflect tolerances, conversion factors, rounding, measurement errors, and other factors known to those skilled in the art, so as to yield equivalent results or effects. In some circumstances, it may not be possible to reasonably determine a value that provides equivalent results or effects. In general, quantities, sizes, formulations, parameters, or other quantities or characteristics are "about," "approximate," or "to, or about," whether or not they are explicitly stated to be so. When "about," "approximate," or "to, or about," is used before a quantitative value, it is understood that the parameter also includes the specific quantitative value itself, unless otherwise specified.

[0111] As used herein, “biological sample” is a sample containing whole cells and / or living cells and / or cellular debris. A biological sample may include (or be derived from) “body fluids.” The present invention encompasses embodiments in which body fluids are selected from amniotic fluid, aqueous humor, vitreous fluid, bile, serum, breast milk, cerebrospinal fluid, earwax (earwax), chyle, atherosclerotic syrup, endolymph, perilymph, exudate, feces, female ejaculate, gastric acid, gastric juice, lymph, mucus (including nasal mucus and sputum), pericardial fluid, ascites, pleural fluid, pus, mucosal secretions, saliva, sebum (skin oil), semen, sputum, synovial fluid, sweat, tears, urine, vaginal secretions, vomit, and mixtures of one or more thereof. Biological samples include cell cultures, body fluids, and cell cultures derived from body fluids. Body fluids can be obtained from mammalian organisms, for example, by puncture or other collection or sampling procedures.

[0112] As used herein, “agent” means any substance, compound, molecule, etc. that can be administered to a subject on which it is administered. An agent may be inert. An agent may be an activator. An agent may be a primary activator, or in other words, a component of a composition to which all or part of the effect of the composition is attributed. An agent may be a secondary agent, or in other words, a component of a composition to which an additional part and / or other effect of the composition is attributed.

[0113] As used herein, “activator” or “active ingredient” means a biologically active or otherwise active substance, compound, or molecule that induces a biological or physiological effect on the subject to which it is administered. In other words, “activator” or “active ingredient” means a component of a composition to which all or part of the effect of the composition is attributable.

[0114] As used herein, “administer” means any appropriate administration of the drug to be delivered and / or for the recipient of the drug, and may be oral, topical, intravenous, subcutaneous, transcutaneous, transdermal, intramuscular, intra-joint, parenteral, arteriole, intradermal, ventricular, intraosseous, intraocular, intracranial, intraperitoneal, intralesional, intranasal, cardiac, intraarticular, intracavitary, intrathecal, intravitreous, intracerebral, intraventricular, intratympanic, cochlear, rectal, vaginal, by inhalation, by catheter, via an implanted reservoir or other device that actively or passively (e.g., by diffusion) administers a stent or composition into the perivascular space and adventitia. For example, a medical device such as a stent may contain a composition or formulation placed on its surface, which can then be dissolved or otherwise distributed into the surrounding tissue and cells. The term "parenteral" may include subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques.The routes of administration include, for example, the auricle (ear), buccal side, conjunctiva, skin, teeth, electroosmosis, intracervical, intranasal sinuses, intratracheal, intestinal, epidural, extraamniotic, extracorporeal, hemodialysis, infiltration, interstitial, intra-abdominal, intraamniotic, intraarterial, intra-articular, intrabile duct, intrabronchial, intrasacral, intracardiac, intracartilage, intrasacral, cavity, intracerebral, cisterna magna, intradental (tooth), and coronary artery. Internal, internal cavernous tissue, intradermal, intravertebral disc, intraductal, duodenal, intradural, intraepidermal, esophageal, gastric, gingival, ileum, lesion, tubular lumen, lymphatic, intramedullary, intrameningeal, intramuscular, intraocular, ovarian, pericardial, intraperitoneal, intrathoracic, prostate, lung, nasal cavity, spinal cord, synovial fluid, tendon, testis, spinal cavity , intrathoracic, intratubal, intratumoral, intratympanic, intrauterine, intravascular, intravenous, intravenous bolus, intravenous infusion, intraventricular, intrabladder, intravitreous, iontophoresis, irrigation, larynx, nasal cavity, nasal cavity-stomach, occlusive bandaging, ophthalmology, oral cavity, oropharynx, other, parenteral, percultaneous, periarticular, peridural, perineurial, periodontal, rectal, respiratory (inhalation), retroocular, soft tissue, subarachnoid, subconjunctival, subcutaneous, sublingual, submucosal, local, transdermal, transmucosal, transplacental, transtracheal, transtympanic, ureter, urethra, and / or vaginal administration, and / or any combination of the above routes of administration, which typically depends on the disease being treated, the subject being treated, and / or the drug being administered.

[0115] As used herein, "cancer" includes acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, Kaposi's sarcoma, AIDS-associated lymphoma, primary central nervous system (CNS) lymphoma, anal cancer, appendiceal cancer, astrocytoma, atypical teratomatoid / rhabdoid tumor, and basal cell tumor of the skin. Cell cancer, bile duct cancer, bladder cancer, bone cancer (including, but not limited to, Ewing's sarcoma, osteosarcoma, and malignant fibrous histiocytoma), brain tumor, breast cancer, bronchial tumor, Burkitt lymphoma, carcinoid tumor, cardiac tumor, germ cell tumor, embryonal tumor, cervical cancer, cholangiocarcinoma, chordoma, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative neoplasm, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, ductal carcinoma in situ, endometrial cancer, ependymoma, esophageal cancer, sensory neuroblastoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer (including, but not limited to, intraocular melanoma and retinoblastoma), fallopian tube cancer, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, central nervous system germ cell tumor Tumors, extracranial germ cell tumors, extragonadal germ cell tumors, ovarian germ cell tumors, testicular cancer, gestational trophoblastic disease, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, Langerhans cell histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer, islet tumors, pancreatic endocrine tumors, renal (renal cell) cancer, laryngeal cancer, leukemia, lip cancer, oral cancer, lung cancer (non-small cell and small cell), lymphoma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous cell carcinoma of the neck, midline cancer with and without NUT gene mutations, multiple endocrine neoplasia syndrome, multiple myeloma, plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, chronic myelogenous leukemia, nasal cavity cancer, sinus cancer cancer), non-Hodgkin lymphoma, pancreatic cancer, paraganglioma, paranasal sinus cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pituitary cancer, peritoneal cancer, prostate cancer, rectal cancer, rhabdomyosarcoma, salivary gland cancer, uterine sarcoma, Sézary syndrome, skin cancer, small intestine cancer, colorectal cancer (colon cancer), soft tissue sarcoma, T-cell lymphoma, pharyngeal cancer (throatThis refers to one or more types of cancer, including but not limited to oropharyngeal cancer, nasopharyngeal cancer, hypopharyngeal cancer, thymoma, thymic cancer, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, urethral cancer, uterine cancer, vaginal cancer, cervical cancer, hemangiomas and cancers, vulvar cancer, and Wilms' tumor.

[0116] As used herein, “chemotherapeutic agent” or “chemotherapeutic drug” refers to a therapeutic agent used to prevent or treat cancer.

[0117] As used herein, “control” is a term of the art and refers to an alternative subject or sample included in an experiment for comparative purposes to minimize or distinguish the effect of a variable other than the independent variable. A control may be positive or negative. The use of an appropriate control to determine a particular effect or the absence of a measurable or observable effect will be understood by those skilled in the art.

[0118] As used herein, the terms “disease” or “disorder” are interchangeable throughout this specification and refer to any change in any condition of any body or organ that interrupts or interferes with the performance of a function and / or causes symptoms such as discomfort, dysfunction, pain, or even death of the person affected or in contact with the person affected. Disease or disorder may also relate to distemper, pathological conditions, illness, disease, disorder, malaise, disease, complaint, malaise, or pain.

[0119] As used herein, “dose,” “unit dose,” or “administered amount” refers to a physically distinct unit suitable for use in the subject, each unit comprising a predetermined amount of Hiroloxamer and / or its pharmaceutical preparation calculated to produce a desired response in relation to its administration.

[0120] As used herein, “Hiroloxamer” refers to a poloxamer 188 (P188) composition that has been purified to have a polydispersity of less than about 1.07 and is free of long-cycled material, as described in U.S. Patent No. 9,403,941.

[0121] The term "molecular weight," as used herein, generally refers to the mass or average mass of a material. In the case of polymers or oligomers, molecular weight may refer to the relative average chain length or relative chain mass of the bulk polymer. In practice, the molecular weight of polymers and oligomers can be estimated or characterized by a variety of methods, including gel permeation chromatography (GPC) or capillary viscosity measurement. GPC molecular weight is the number average molecular weight (M n In contrast to ), weight-average molecular weight (M w It is reported as follows: Capillary viscosity measurement provides an estimate of molecular weight as intrinsic viscosity determined from a diluted polymer solution using a specific set of concentration, temperature, and solvent conditions.

[0122] The term "depending on the circumstances" means that the event, situation, or substituent described thereafter may or may not occur, and that the description includes both cases in which the event or situation occurs and cases in which it does not.

[0123] As used herein, “pharmaceutical preparation” refers to a combination of an activator, compound, or component that makes the composition suitable for in vitro, in vivo, or ex vivo diagnostic, therapeutic, or prophylactic use, and a pharmaceutically acceptable carrier or excipient.

[0124] As used herein, “pharmaceutically acceptable carrier or excipient” means a carrier or excipient that is generally safe, non-toxic, and useful for preparing pharmaceutical formulations that are not biologically or otherwise undesirable, and includes carriers or excipients that are acceptable for veterinary and human pharmaceutically use. As used herein and in the claims, “pharmaceutically acceptable carrier or excipient” includes both one and two or more such carriers or excipients.

[0125] As used herein, “polymer” refers to a molecule composed of linked monomer repeating units. “Polymer” is understood to include, but is not limited to, homopolymers, copolymers, e.g., block, graft, random, and alternating copolymers, terpolymers, and blends and modifications thereof. “Polymer” can be a three-dimensional network (e.g., repeating units linked together left and right, front and back, top and bottom), a two-dimensional network (e.g., repeating units in sheet form linked together left, right, top and bottom), or a one-dimensional network (e.g., repeating units linked left and right to form a chain). “Polymer” can be composed of natural monomers or synthetic monomers and combinations thereof. Polymers may be biological (e.g., monomers may be biologically important (e.g., amino acids)), natural, or synthetic.

[0126] As used herein, "preventive," "preventive," "preventive," and "prophylactic" refer to interfering with or cessating a disease or condition before it occurs (by the action of a compound, formulation, and / or method), even if it has not been diagnosed or while the disease or condition is still in the subclinical stage.

[0127] As used herein, the term “radiosensitizer” refers to an agent that can selectively enhance cell death from irradiation in a desired cell population, such as tumor cells, without exhibiting monotoxicity to tumor cells or normal cells.

[0128] The terms “subject,” “individual,” and “patient” are used interchangeably herein to refer to vertebrates, preferably mammals, more preferably humans. Mammals include, but are not limited to, mice, monkeys, humans, livestock, sports animals, and pets. Tissues, cells, and their offspring of biological entities obtained in vivo or cultured in vitro are also included in the term “subject.”

[0129] As used herein, “substantially pure” means the species of interest, which is the dominant species present (i.e., more abundant in molar terms than any other individual species in the composition), and preferably, the substantially purified fraction is a composition in which the species of interest constitutes about 50 percent of all species present. Generally, a substantially pure composition will consist of more than about 80 percent, more preferably more than about 85%, 90%, 95%, and 99 percent of all species present in the composition. Most preferably, the species of interest is purified to an intrinsically homogeneous state in which the composition consists essentially of a single species (conventional detection methods cannot detect any contaminating species in the composition).

[0130] Where used interchangeably in this specification, the terms “sufficient” and “effective” refer to the amount (e.g., mass, volume, dose, concentration, and / or duration) required to achieve one or more desired and / or described results. For example, a therapeutically effective dose refers to the amount required to achieve one or more therapeutic effects.

[0131] As used herein, “tangible medium” means a medium that is physically tangible or accessible and is not merely abstract thought or unrecorded spoken words. “Tangible medium” includes, but is not limited to, words on cellulose or plastic material, or data stored in a suitable computer-readable memory form. The data may be stored on a unit device such as flash memory or a CD-ROM, or on a server that can be accessed by a user, for example, via a web interface.

[0132] As used herein, “therapeutic” means treating, curing, and / or improving a disease, disorder, condition, or side effect, or slowing the progression of a disease, disorder, condition, or side effect. Accordingly, “therapeutic dose” may refer to the amount of compound that can produce a therapeutic effect.

[0133] As used herein, the terms “to treat” and “treatment” generally refer to obtaining a desired pharmacological and / or physiological effect. The effect may be, but not necessarily, prevent or partially prevent a disease, symptom, or condition, such as cancer and / or indirect radiation injury. The effect may be therapeutic in the sense of partial or complete cure of a disease, condition, symptom, or adverse effect resulting from a disease, disorder, or condition. As used herein, the term “treatment” encompasses any treatment of cancer and / or indirect radiation injury in a subject, particularly in humans and / or companion animals, and may include any one or more of the following: (a) preventing the development of a disease or injury in a subject that may be predisposed to the disease but has not yet been diagnosed with it; (b) inhibiting the disease, i.e., stopping its onset; and (c) alleviating the disease, i.e., alleviating or improving the disease and / or its symptoms or condition. As used herein, the term “treatment” may refer to a therapeutic treatment alone, a prophylactic (preventive) treatment alone, or both a therapeutic and a prophylactic treatment. Those who require treatment (the subjects requiring it) may include those who already have a disability and / or those for whom disability should be prevented. As used herein, the term “to treat” may include inhibiting a disease, disability, or condition, e.g., preventing its progression; and alleviating a disease, disability, or condition, e.g., causing regression of the disease, disability, and / or condition. Treating a disease, disability, or condition may include improving at least one symptom of a particular disease, disability, or condition, even if the underlying pathophysiology is not affected, e.g., treating pain in a subject by administering an analgesic, even if the analgesic does not treat the cause of the pain.

[0134] Where used herein, the terms “weight percentage,” “wt%,” and “wt%” refer to the weight percentage of a given component based on the total weight of the composition in which it is a component, unless otherwise specified. That is, unless otherwise specified, all weight percentage values ​​are based on the total weight of the composition. It should be understood that the sum of the weight percentage values ​​of all components in a disclosed composition or formulation equals 100. Alternatively, if the weight percentage values ​​are based on the total weight of a subset of components in a composition, it should be understood that the sum of the weight percentage values ​​of a particular component in the disclosed composition or formulation equals 100.

[0135] As used herein, "water-soluble" generally means that at least about 10 g of a substance is soluble in 1 L of water, i.e., at a neutral pH, at 25°C.

[0136] Various embodiments are described below. It should be noted that no particular embodiment is intended to be exhaustive or to be a limitation to a broader set of embodiments discussed herein. An embodiment described in relation to a particular embodiment is not necessarily limited to that embodiment and can be implemented in any other embodiment. Throughout this specification, the references “one embodiment,” “embodiment,” and “exemplary embodiment” mean that a particular feature, configuration, or characteristic described in relation to an embodiment is included in at least one embodiment of the present invention. Thus, occurrences of the phrases “in one embodiment,” “in an embodiment,” or “in an exemplary embodiment” in various places throughout this specification do not necessarily all refer to the same embodiment, although they may refer to the same embodiment. Furthermore, certain features, structures, or characteristics can be combined in any suitable manner in one or more embodiments, as will be apparent to those skilled in the art from this disclosure. Furthermore, while some embodiments described herein include some features included in other embodiments but not others, combinations of features from different embodiments are intended to be within the scope of the present invention. For example, in the appended claims, any of the claimed embodiments can be used in any combination.

[0137] All publications, published patent documents, and patent applications cited herein are incorporated herein by reference to the same extent as each individual publication, published patent document, or patent application is specifically and individually indicated as being incorporated by reference.

[0138] overview Radiotherapy is a well-established modality for cancer treatment and has been used for over 50 years. Radiation is also involved in other fields such as nuclear energy and research. Therefore, radiation exposure can be both intentional (e.g., for therapeutic purposes) and accidental or unintentional (e.g., radiation leakage or exposure to a contaminated environment). Radiation (regardless of the source) directly causes DNA damage to cells, such as single-strand breaks (SSBs), double-strand breaks (DSBs), DNA crosslinks, and DNA-protein crosslinks, or indirectly induces DNA damage by reactive oxygen species (ROS) / reactive nitrogen species (RNS), which leads to cellular senescence and apoptosis. Radiation, in its essence, does not distinguish between cancerous and non-cancerous cells. While methods and techniques have been developed to target radiation only to cancerous or specific types of cells or tissues in therapeutic contexts (e.g., sensitizing cancerous cells, thereby requiring less radiation and physically limiting the area of ​​direct exposure), radiotherapy is still considered a "double-edged sword" and can still cause significant damage to non-cancerous cells. Furthermore, accidental radiation exposure presents a significant health risk because, in most cases, precautionary measures to limit exposure are often not taken or are insufficient, as if the exposure were not planned. Other injuries, such as mechanical, chemical (e.g., hydrogen peroxide or other drugs), or biological (e.g., viruses and microorganisms), can result in ROS, RNS, peroxides, superoxides, or other inflammatory damage in surrounding, and sometimes distant, cells and tissues that are not directly exposed to the injury.

[0139] Despite the enormous benefits of radiotherapy and diagnostics, ionizing and other types of radiation can be harmful to otherwise normal and healthy cells. See, for example, Desouky et al., 2015, J. Rad Res App Sci. 8(2):247-254. Ionizing radiation is energetic and penetrating and has direct and indirect effects. Directly, radiation directly collides with DNA molecules, altering their molecular structure, which can lead to cell damage and even cell death. Surviving cells with damage may later become cancerous or have other abnormalities that impair normal function. Indirectly, radiation collides with water molecules and other organic molecules inside and outside cells, thereby generating free radicals such as hydroxyl and alkoxy moieties. Reactive nitrogen species are also generated by the direct ionization of DNA, which can also have harmful indirect effects.

[0140] Furthermore, radiation can induce what is known in this art as the "bystander effect." Ionizing radiation-induced bystander effects can be defined as biological effects on unirradiated cells resulting from the exposure of other cells in a population to radiation. These bystander effects are more pronounced with low doses of radiation. Therefore, current efforts to make radiotherapy safer by using less radiation may actually increase the incidence of harmful bystander effects in healthy, non-cancerous cells. Bystander effects are thought to be due to bystander signals that can be transmitted via direct cell-to-cell contact or via soluble factors released into the environment and circulation (e.g., reactive oxygen species and reactive nitrogen species, cytokines, calcium ions, and small RNAs) (see, e.g., JBLittle.2006, Mut.Res.Fund.Molc.Mech.Mutagen.597:113-118). See also Williams, JPMcBride, WH, 2011. Int J Radiat Biol 87(8):851-868.

[0141] Similarly, though not bound by theory, the “bystander effect” observed in radiation exposure situations may also occur secondarily to mechanical, chemical, and / or biological damage resulting in the production of ROS, RNS, peroxides, superoxides, and / or inflammatory responses. Therefore, there is still a need for compositions, methods, and techniques to protect cells and tissues that are not directly exposed to damage resulting in the production of ROS, RNS, peroxides, superoxides, or other inflammatory damage in surrounding, and sometimes distant, cells and tissues.

[0142] Accordingly, embodiments disclosed herein can provide methods for protecting uninjured cells during and / or after exposure to injury. As used herein, “uninjured cells” refer to cells adjacent to and / or distant to injured cells that have not been directly exposed to injury, which may be mechanical, chemical, biological, or energetic. It will be understood that injury causes negative changes and / or damage to cells it directly affects. Injury may result in the production of free radicals (e.g., ROS, RNS, peroxides, superoxides), as well as inflammation from injured and / or uninjured cells. As noted above, adjacent and distant cells may still be indirectly damaged by “bystander effects” among other mechanisms. In some embodiments, the method may include administering a first amount of poloxamer 188 (P188) or a pharmaceutical formulation thereof to one or more areas of the subject before exposing them to injury; and exposing one or more areas of the subject to injury (intentionally (for treatment) or accidentally) after the blood concentration of P188 has reached approximately 1 to 5 mg / mL, for example, approximately 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or approximately 5 mg / mL.

[0143] In some embodiments, the method may include administering a first amount of poloxamer 188 (P188) or a pharmaceutical formulation thereof to a subject about 0 hours (i.e., at the time of injury) to 48 hours before exposing one or more areas of the subject to injury; and exposing one or more areas of the subject to injury (either intentionally (e.g., for treatment) or accidentally).

[0144] Also provided herein are kits that include an effective dose of P188 or a pharmaceutical formulation thereof, and instructions for administration 0 to 48 hours before exposure of one or more areas of a subject to ionizing radiation. In some embodiments, the method can prevent cellular damage in proximal and / or distal to injured cells, or in a larger number of uninjured cells, in a subject exposed to injury.

[0145] Embodiments disclosed herein can provide methods for protecting cells, particularly non-cancerous cells and / or non-irradiated cells, during and / or after radiation exposure, such as that occurring during radiotherapy for cancer. In some embodiments, the method may include administering a first dose of poloxamer 188 (P188) or a pharmaceutical formulation thereof to one or more areas of the area before exposing them to ionizing radiation; and exposing one or more areas of the area to ionizing radiation after the blood concentration of P188 has reached approximately 1 to 5 mg / mL, for example, approximately 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or approximately 5 mg / mL.

[0146] In some embodiments, the method may include administering a first dose of poloxamer 188 (P188) or a pharmaceutical formulation thereof to a subject about 0 to 48 hours before exposure of one or more areas of the subject to ionizing radiation; and exposing one or more areas of the subject to ionizing radiation. Also provided herein are kits that include an effective dose of P188 or a pharmaceutical formulation thereof, and instructions for administration 0 to 48 hours before exposure of one or more areas of the subject to ionizing radiation. In some embodiments, the method may prevent cellular damage, such as non-cancerous and / or non-irradiated cell damage, in a subject exposed to ionizing radiation.

[0147] While not bound by theory, the poloxamer 188 formulations described herein can coat cells, as opposed to being sequestered in the blood, and can provide protection from free radicals, DAMPs, and components of the inflammatory cascade. Free radicals, DAMPs, and components of the inflammatory cascade may be the result of or products generated from injured cells adjacent to or distant from uninjured cells, and can cause damage to uninjured cells. The compositions and methods described herein may have the advantages of protecting uninjured cells, repairing injured cells, and / or mitigating injury or inflammatory processes, for example, by preserving cells and / or tissues that would otherwise be lost due to injury and various downstream effects. See also Goliaei et al., 2017. J Phys Chem B. 120(33) 8361-8641. Furthermore, embodiments of the P188 formulations described herein are thought to involve hydrophobic-hydrophilic association interactions with cell surface lipid bilayer membranes. When placed on the cell surface, the polymer forms a network of intermolecular hydrogen bonds between polymer molecules and between polymer molecules and water. This protective layer can capture and quench free radicals and close damage-related pores in the cell membrane by increasing the packing density of the lipid bilayer, resulting in one or more effects described herein. This may enable cells to absorb and / or neutralize free radicals. This can close holes in the cell membrane and repair damaged areas of the cell membrane. This can minimize the release of inflammatory molecules and damage-related molecular patterns. This can also provide blood rheological modifications that may be relevant to erythrocytes and vascular endothelium, reducing viscosity and cell-cell interactions. In some embodiments, purified forms of P188 (including, but not limited to, Hiroloxamer) can result in these specific mechanisms of action.

[0148]

[0149] Other compositions, compounds, methods, features, and advantages of this disclosure will be apparent to those skilled in the art upon consideration of the following drawings, detailed description, and examples. All such additional compositions, compounds, methods, features, and advantages are contained herein and are intended to be within the scope of this disclosure.

[0150] Poloxamer preparations Pharmaceutical formulations are described herein that may contain a certain amount, effective amount, and / or minimum effective amount, and / or therapeutically effective amount of a suitable poloxamer, such as P188 (also referred to elsewhere herein as a primary activator or component), and a pharmaceutically acceptable carrier. The P188 pharmaceutical formulation may then be administered to a subject requiring it in accordance with the methods described elsewhere herein.

[0151] Poloxamer is a synthetic triblock copolymer consisting of two hydrophilic polyoxyethylene chains adjacent to a central hydrophobic polyoxypropylene chain, having a weight ratio of 4:2:4, and conforming to formula I (where a=80 and b=27). [ka]

[0152] This configuration results in amphiphilic surface copolymers in which molecular size, hydrophilicity, and hydrophobicity can be varied by changing the number of central and side chains of the molecule. Poloxamers are generally abbreviated with the letter "P" (in the case of poloxamers) and the following three digits. Multiplying the first two digits by 100 gives the approximate molecular mass of the polyoxypropylene core, and multiplying the last digit by 10 gives the percentage of polyoxyethylene content. Poloxamer 188 (P188) is a nonionic linear copolymer with an average molecular weight of 8400 Daltons. It is commercially known as PLURONIC F68, FLOCOR, and RheothRx. P188 is also described in U.S. Patent No. 5,696,298.

[0153] In some embodiments, P188 can be chemically synthesized via a two-step process. In the first step, a (poly)oxypropylene core is constructed, and in the second step, poly(oxyethylene) is added to the ends of the polyoxypropylene core. Due to variations in polymerization rates between both steps, the synthesized P188 contains a bell-shaped distribution of polymer species, mainly with variations in total chain length. In some embodiments, P188 can be filtered to reduce its polydispersity to produce purified P188 suitable for use in embodiments of the method described herein. In some embodiments, any present low molecular weight products are removed. In some embodiments, both present low molecular weight and high molecular weight products are removed by any suitable size fractionation or separation technique.

[0154] In some embodiments, the pharmaceutical formulation contains a certain amount of purified P188 such that the P188 formulation is substantially free of low molecular weight products (i.e., products having a molecular weight of about 5,500 Da or less). In some embodiments, the formulation contains purified P188 such that the formulation is substantially free of low molecular weight products and substantially free of high molecular weight products (i.e., products having a molecular weight of about 16,000 Da or more). In some embodiments, the P188 formulations described herein contain P188 molecules having molecular weights in the range of 7,000 Da to about 15,500 Da. In some embodiments, the average molecular weight of the P188 molecules is about 8,740. In some embodiments, the average molecular weight of P188 is about 8,740, and the molecular size is in the range of about 7,000 Da to about 15,500 Da. In some embodiments, the polydispersity is about 1.07 or less, for example (1.06, 1.05, 1.04, 1.03, 1.02, 1.01, or 1).

[0155] In some embodiments, P188 is Hiroloxamer. In some embodiments, P188 is the P188 composition described in U.S. Patent No. 9,403,941.

[0156] In some embodiments, the pharmaceutical formulation contains a certain amount of Hiroloxamer.

[0157] Pharmaceutically acceptable carriers, auxiliary components, and drugs Pharmaceutical formulations may include pharmaceutically acceptable carriers. Suitable pharmaceutically acceptable carriers include, but are not limited to, water, salt solutions, alcohols, gum arabic, vegetable oils, benzyl ethyl alcohol, polyethylene glycol, gelatin, carbohydrates such as lactose, amylose or starch, magnesium stearate, talc, silicic acid, viscous paraffin, fragrance oils, fatty acid esters, hydroxymethylcellulose, and polyvinylpyrrolidone, which do not react adversely with the active composition.

[0158] Pharmaceutical preparations can be sterilized and, if desired, can be mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts to affect osmotic pressure, buffers, colorants, flavoring agents and / or aromatic substances, which do not react harmfully with the active compound.

[0159] In some embodiments, the pharmaceutical formulation may also contain an effective amount of co-activators, including but not limited to biological agents or molecules (e.g., polypeptides, polynucleotides, antibodies and their fragments, aptamers, etc.), chemotherapeutic agents, antineoplastic agents, hormones, antibiotics, antiviral agents, immunomodulators, antiemetics, pain-modifying compounds (such as opiates), anti-inflammatory agents, antipyretics, antibiotics, and combinations thereof.

[0160] Effective amount In some embodiments, the amount of the primary activator (e.g., P188 or purified P188) and / or optional auxiliary activators may be an effective amount, a minimum effective amount, and / or a therapeutically effective amount. The effective amount, minimum effective amount, and / or therapeutically effective amount of the primary activator and / or auxiliary activator, as used in other parts of this specification, contained in the pharmaceutical preparation, is approximately 0 or approximately 1 to 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490. , 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, or a non-zero amount in any range of ~1000 pg, ng, μg, mg, or g, or any numerical range having any of these ranges.In some embodiments, the effective dose, minimum effective dose, and / or therapeutically effective dose may be the effective concentration, minimum effective concentration, and / or therapeutically effective concentration, each of which is approximately 0 or approximately 1-10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 4 It may be a non-zero quantity in the range of any number having any of the following ranges: 90, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, or ~1000pM, nM, μM, mM, or M.

[0161] In other embodiments, the effective amount, minimum effective amount, and / or therapeutically effective amount of the auxiliary activator present in the formulation is approximately 0 or approximately 1 to 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500. , 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, or ~1000 IU, or any range of numbers having any of these ranges, which may be non-zero quantities.

[0162] In some embodiments, the primary activator is present in the pharmaceutical formulation at approximately 0-0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.3% of the pharmaceutical formulation. , 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, 0.5, 0.51, 0.52, 0.53, 0.54, 0.55, 0.56, 0.57, 0.58, 0.59, 0.6, 0.61, 0.62, 0.63, 0.64, 0.65, 0.66, 0.67, 0.68, 0.69, 0.7, 0.71, 0.72, 0.73, 0.74, 0.75, 0.76, 0.77, 0.78 ,0.79,0.8,0.81,0.82,0.83,0.84,0.85,0.86,0.87,0.88,0.89,0.9,0.91,0.92,0.93,0.94,0.95,0.96,0.97,0.98,0.9,1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,4 It may be a non-zero quantity in the range of 8, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, 99.9%, or ~approximately 100% w / w, v / v, or w / v.

[0163] In some embodiments, if present, the auxiliary activator is present in amounts of approximately 0-0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29 of the pharmaceutical formulation. 0.3, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, 0.5, 0.51, 0.52, 0.53, 0.54, 0.55, 0.56, 0.57, 0.58, 0.59, 0.6, 0.61, 0.62, 0.63, 0.64, 0.65, 0.66, 0.67, 0.68, 0.69, 0.7, 0.71, 0.72, 0.73, 0.74, 0.75, 0.76, 0.77, 0. 78, 0.79, 0.8, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.9, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, These can be non-zero quantities in the range of 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, 99.9%, or ~approximately 100% w / w, v / v, or w / v.

[0164] In embodiments in which a co-activating agent is included in the pharmaceutical formulation, the effective amount of the co-activating agent will vary depending on the co-activating agent.

[0165] If present in a pharmaceutical preparation, the auxiliary activator may be contained in the pharmaceutical preparation, or it may exist as a compound, a derivative thereof, or as an independent compound or pharmaceutical preparation that can be administered simultaneously or sequentially with the pharmaceutical preparation. In other embodiments, the effective amount of the auxiliary activator is approximately 0-1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58 of the total auxiliary activator preparation. , 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, ~ may be non-zero quantities in the range of w / w, v / v, or w / v. In additional embodiments, the effective amount of the auxiliary activator is approximately 0-1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 of the total formulation. 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, or non-zero quantities in the range of ~99.9% w / w, v / v, or w / v.

[0166] Dosage form In some embodiments, the pharmaceutical formulations described herein may be dosage forms. A dosage form can be administered to a target requiring it. A dosage form can effectively generate a specific concentration, such as an effective concentration, at a given site in the target requiring it. In some cases, a dosage form contains a larger amount of the active ingredient or co-active ingredient than the final intended amount required to reach a specific region or location within the target.

[0167] The dosage form can be adapted for administration via any suitable route. Suitable routes include, but are not limited to, oral (including buccal or sublingual), rectal, intraocular, inhalation, intranasal, topical (including buccal, sublingual, or transdermal), vaginal, parenteral, subcutaneous, intramuscular, intravenous, intranasal, and intradermal. Other suitable routes are described elsewhere in this specification. Such formulations can be prepared by any method known in the art.

[0168] Dosage forms suitable for oral administration may be capsules, pellets or tablets, powders or granules, liquids, or suspensions in aqueous or non-aqueous liquids; individual dosage forms such as edible foams or whips, or oil-in-water or water-in-oil liquid emulsions. In some embodiments, a pharmaceutical preparation suitable for oral administration may also include one or more agents that help flavor, preserve, color, or disperse the pharmaceutical preparation. Dosage forms prepared for oral administration may also be in the form of liquid solutions that can be delivered as foams, sprays, or liquid solutions. Oral dosage forms can be administered to subjects requiring them. Where appropriate, the dosage forms described herein may be microencapsulated.

[0169] Dosage forms can also be prepared to extend or prolong the release of any component. In some embodiments, compounds, molecules, compositions, vectors, vector systems, cells, or combinations thereof described herein may be components whose release is delayed. In some embodiments, the primary activator is the component whose release is delayed. In some embodiments, an optional auxiliary agent may be the component whose release is delayed. Suitable methods for delaying the release of a component include, but are not limited to, coating or embedding the component in a material such as a polymer, wax, or gel. Delayed-release formulations can be prepared as described in standard references such as “Pharmaceutical dosage form tablets,” eds. Liberman et al. (New York, Marcel Dekker, Inc., 1989), “Remington—The science and practice of pharmacy,” 20th ed., Lippincott Williams & Wlkins, Baltimore, MD, 2000, and “Pharmaceutical dosage forms and drug delivery systems,” 6th Edition, Ansel et al., (Media, PA: Williams and Wlkins, 1995). These references provide information on excipients, materials, apparatus, and processes for preparing tablets and capsules, as well as information on delayed-release dosage forms of tablets and pellets, capsules, and granules. Delayed release may range from about 1 hour to about 3 months or longer.

[0170] Examples of suitable coating materials include, but are not limited to, cellulose polymers such as cellulose acetate phthalate, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and hydroxypropyl methylcellulose acetate succinate; polyvinyl acetate phthalate, acrylic acid polymers and copolymers, and methacrylic resins, zein, shellac, and polysaccharides, which are commercially available under the trade name EUDRAGIT® (Roth Pharma, Westerstadt, Germany).

[0171] The coating can be formed with or without water-insoluble / water-soluble nonpolymer excipients in different ratios of water-soluble polymers, water-insoluble polymers, and / or pH-dependent polymers to produce a desired release profile. The coating is performed on a dosage form (matrix or simple) containing “as-is” components that are formulated as tablets (compressed with or without coated beads), capsules (with or without coated beads), beads, particulate compositions, suspensions, or sprinkle dosage forms, but is not limited to these.

[0172] Where appropriate, the dosage forms described herein may be liposomes. In these embodiments, the primary active ingredient, and / or optionally a co-active ingredient, and / or, where appropriate, a pharmaceutically acceptable salt thereof, is incorporated into the liposome. In embodiments where the dosage form is a liposome, the pharmaceutical formulation is therefore a liposomal formulation. The liposomal formulation can be administered to a subject requiring it. Dosage forms suitable for topical administration can be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols, or oils. In some embodiments for the treatment of the eye or other external tissue, such as the mouth or skin, the pharmaceutical formulation is applied as a topical ointment or cream. When formulated as an ointment, the primary active ingredient, optionally a co-active ingredient, and / or, where appropriate, a pharmaceutically acceptable salt thereof may be formulated with a paraffinic or water-miscible ointment base. In other embodiments, the primary and / or co-active ingredients may be formulated in a cream with an oil-in-water or water-in-oil base. Suitable dosage forms for local administration in the mouth include lozenges, troches, and mouthwashes.

[0173] Dosage forms suitable for nasal or inhalation administration include aerosols, solutions, suspensions, gels, or dry powders. In some embodiments, the primary active ingredient, optionally co-active ingredients, and / or pharmaceutically acceptable salts thereof may, where appropriate, be in a dosage form suitable for inhalation and may be obtained or be obtained by micronization, resulting in a reduced particle size. In some embodiments, the reduced particle size (e.g., micronized) of the compound or its salt or solvate is measured by a suitable method known in the art to be about 0.5 to about 10 microns. 50 This is determined by the value. Dosage forms suitable for inhalation administration also include particulate dust or mist. Suitable dosage forms in which the carrier or excipient is a liquid for administration as a nasal spray or nasal drop include aqueous or oily solutions / suspensions of the active (primary and / or auxiliary) component, which can be produced by various types of metered-dose pressurized aerosols, nebulizers, or inhalers. Nasal / inhalation formulations can be administered to subjects who require them.

[0174] In some embodiments, the dosage form is an aerosol formulation suitable for administration by inhalation. In some of these embodiments, the aerosol formulation comprises, where appropriate, a solution or fine suspension of a primary active ingredient, a co-active ingredient, and / or a pharmaceutically acceptable salt thereof, in a pharmaceutically acceptable aqueous or non-aqueous solvent. The aerosol formulation may exist in a single or multi-dose form in a sealed container. In some of these embodiments, the sealed container is a single-dose or multi-dose nasal or aerosol dispenser equipped with a metering valve (e.g., a metering inhaler), intended to be disposed of once the contents of the container have been used up.

[0175] When the aerosol formulation is contained in an aerosol dispenser, the dispenser contains a suitable propellant under pressure, such as compressed air, carbon dioxide, or an organic propellant, including but not limited to hydrofluorocarbons. In other embodiments, the aerosol formulation formulation is contained in a pump atomizer. The pressurized aerosol formulation may also contain a solution or suspension of a primary active ingredient, optionally a co-active ingredient, and / or a pharmaceutically acceptable salt thereof. In further embodiments, the aerosol formulation may also contain a co-solvent and / or modifier incorporated to improve, for example, the stability and / or taste and / or particulate mass characteristics (quantity and / or profile) of the formulation. The aerosol formulation may be administered once or several times a day, for example, two, three, four, or eight times a day, with one, two, or three doses delivered each time. The aerosol formulation may be administered to a subject that requires it.

[0176] For certain dosage forms suitable and / or adapted for inhalation administration, the pharmaceutical formulation is a dry powder inhalable formulation. Where appropriate, in addition to a primary activator, optionally co-active ingredients, and / or pharmaceutically acceptable salts thereof, such a dosage form may include a powder base such as lactose, glucose, trehalose, mannitol, and / or starch. In some of these embodiments, the primary activator, co-active ingredients, and / or pharmaceutically acceptable salts thereof are, where appropriate, in a reduced particle size form. In further embodiments, performance modifiers, such as L-leucine or another amino acid, cellobiose octaacetate, and / or metal salts of stearic acid, such as magnesium stearate or calcium stearate. In some embodiments, the aerosol formulation is prepared such that each quantitative aerosol contains a predetermined amount of an active ingredient, such as one or more of the compositions, compounds, vectors, molecules, cells, and combinations thereof described herein.

[0177] Dosage forms suitable for vaginal administration may exist as pessaries, tampons, creams, gels, pastes, foams, or sprays. Dosage forms suitable for rectal administration include suppositories or enemas. Vaginal preparations can be administered to those who require them.

[0178] Dosage forms suitable for parenteral administration and / or injection may include aqueous and / or non-aqueous sterile injection solutions that may contain antioxidants, buffers, bacteriostatic agents, and solutes that make the composition isotonic with the target blood, as well as aqueous and non-aqueous sterile suspensions that may contain suspending agents and thickeners. Dosage forms suitable for parenteral administration may exist in single-unit or multi-unit dose containers, including but not limited to sealed ampoules or vials. Dosages may be lyophilized and resuspended on a sterile carrier to reconstitute the dose before administration. Immediate injection solutions and suspensions may, in some embodiments, be prepared from sterile powders, granules, and tablets. Parenteral formulations may be administered to subjects requiring them.

[0179] In some embodiments, the dosage form comprises, where appropriate, a predetermined amount of a primary activator, a co-active ingredient, and / or a pharmaceutically acceptable salt thereof per unit dose. In one embodiment, the predetermined amount of the primary activator, the co-active ingredient, and / or a pharmaceutically acceptable salt thereof may, where appropriate, be an effective dose, a minimum effective dose, and / or a therapeutically effective dose. In some embodiments, the predetermined amount may be effective in protecting unirradiated cells when one or more areas of a subject are exposed to ionizing radiation.

[0180] In other embodiments, a predetermined amount of a primary activator, an auxiliary activator, and / or pharmaceutically acceptable salts thereof may, where appropriate, be a suitable fraction of an effective amount of the active ingredient. Thus, such a unit dose may be administered once or more times a day, month, or year (e.g., once, two, three, four, five, six times, or more times a day, month, or year). Such pharmaceutical formulations may be prepared by any method well known in the art.

[0181] When co-therapies, adjuvants, and / or multiple pharmaceutical formulations are delivered to a target and / or cells, different therapies or formulations may be administered sequentially or simultaneously. Sequential administration refers to administrations where a significant amount of time elapses between doses, such as approximately 15, 20, 30, 45, 60 minutes or more. The time between doses in sequential administration can be on the order of hours, days, months, or even years, depending on the active ingredients present in each dose. Simultaneous administration refers to administering two or more formulations simultaneously, or substantially simultaneously (e.g., within seconds or minutes apart), with the intention being to administer the formulations at the same time. In some embodiments, adjuvants, co-therapies, etc., may be administered before, after, or both before and after the poloxamer formulation.

[0182] Exemplary adjuvant activator / co-therapy In some of the embodiments described above, one or more additional compounds may be administered as co-activators. Such co-activators may include, but are not limited to, polynucleotides, amino acids, peptides, polypeptides, antibodies, aptamers, ribozymes, essential oncoproteins and guide sequences for ribozymes that inhibit the translation or transcription of genes, hormones, immunomodulators, antipyretics, anxiolytics, antipsychotics, analgesics, antispasmodics, anti-inflammatory drugs, antihistamines, anti-infective drugs, chemotherapeutic agents, radiosensitizers, and combinations thereof.

[0183] Suitable radiosensitizers include, but are not limited to, 5-fluorouracil, platinum analogs (e.g., cisplatin, carboplatin, and oxaliplatin), gemcitabine, DNA topoisomerase I targeters (e.g., camptothecin derivatives (e.g., topotecan and irinotecan)), epidermal growth factor receptor blockers (e.g., cetuximab, gefitinib), farnesyltransferase inhibitors (e.g., L-778-123), COX-2 inhibitors (e.g., rofecoxib, celecoxib, and etoricoxib), bFGF and VEGF targeters (e.g., bevazucimab and thalidomide), NBTXR3, Nimoral, transcrocetinate sodium, NVX-108, and combinations thereof. For example, see Kvols, LK., J Nucl Med 2005;46:187S-190S.

[0184] Appropriate hormones include, but are not limited to, amino acid-derived hormones (e.g., melatonin and thyroxine), small peptide hormones and protein hormones (e.g., thyroid-stimulating hormone-releasing hormone, vasopressin, insulin, growth hormone, luteinizing hormone, follicle-stimulating hormone, and thyroid-stimulating hormone), eicosanoids (e.g., arachidonic acid, lipoxin, and prostaglandins), and steroid hormones (e.g., estradiol, testosterone, tetrahydrotestosterone, and cortisol).

[0185] Appropriate immunomodulatory agents include, but are not limited to, prednisone, azathioprine, 6-MP, cyclosporine, tacrolimus, methotrexate, interleukins (e.g., IL-2, IL-7, and IL-12), cytokines (e.g., interferons (e.g., IFN-α, IFN-β, IFN-ε, IFN-K, IFN-ω, and IFN-γ), granulocyte colony-stimulating factor, and imiquimod), chemokines (e.g., CCL3, CCL26, and CXCL7), cytosine phosphate-guanosine, oligodeoxynucleotides, glucans, antibodies, and aptamers).

[0186] Appropriate antipyretics include, but are not limited to, nonsteroidal anti-inflammatory drugs (e.g., ibuprofen, naproxen, ketoprofen, and nimeslide), aspirin and related salicylates (e.g., choline salicylate, magnesium salicylate, and sodium salicylate), paracetamol / acetaminophen, metamizole, nabumetone, phenazone, and quinine.

[0187] Appropriate anxiolytics include, but are not limited to, benzodiazepines (e.g., alprazolam, bromazepam, chlordiazepoxide, clonazepam, clorazepate, diazepam, flurazepam, lorazepam, oxazepam, temazepam, triazolam, and tofisopam), serotonergic antidepressants (e.g., selective serotonin reuptake inhibitors, tricyclic antidepressants, and monoamine oxidase inhibitors), Mevicar, favomotisol, selanc, bromantan, emoxipine, azapirone, barbiturates, hydroxyzine, pregabalin, validol, and beta-blockers.

[0188] Appropriate antipsychotic medications include bemperidol, bromoperidol, droperidol, haloperidol, moperone, pipemperone, timiperone, fluspirylene, penfluridol, pimozide, acepromazine, chlorpromazine, siamemazine, dixilazine, fluphenazine, levomepromazine, mesolidazine, perazine, periciazine, perphenazine, pipothiazine, prochlorperazine, promazine, promethazine, protipendyl, thioproperazine, thioridazine, trifluoperazine, triflupromazine, chlorprothixen, clopentixol, flupentixol, thiothixen, zuclopentixol, clotiapine, roxapine, and protipe Examples include, but are not limited to, diclonapine, carpipramine, clocapramine, morindone, mosapramine, sulpiride, belaripride, amisulpride, amoxapine, aripiprazole, asenapine, clozapine, blonanserin, iloperidone, lurasidone, merperone, nemonapride, olanzapine, paliperidone, perospirone, quetiapine, remoxipridone, risperidone, certindol, trimipramine, ziprasidone, zotepine, alstony, bifepurnox, vitopertin, brexpiprazole, cannabidiol, caliprazine, pimavanserin, pomaguritadomethionyl, babicaserin, xanomeline, and diclonapine.

[0189] Appropriate analgesics include, but are not limited to, paracetamol / acetaminophen, nonsteroidal anti-inflammatory drugs (e.g., ibuprofen, naproxen, ketoprofen, and nimeslide), COX-2 inhibitors (e.g., rofecoxib, celecoxib, and etoricoxib), opioids (e.g., morphine, codeine, oxycodone, hydrocodone, dihydromorphine, pethidine, buprenorphine), tramadol, norepinephrine, flupirin, nehopam, orphenadrine, pregabalin, gabapentin, cyclobenzaprine, scopolamine, methadone, ketobemidone, pyritramide, and aspirin and related salicylates (e.g., choline salicylate, magnesium salicylate, and sodium salicylate).

[0190] Appropriate antispasmodics include, but are not limited to, mebeverine, papaverine, cyclobenzaprine, carisoprodol, orphenadrine, tizanidine, metaxalone, methocarbamol, chlorzoxazone, baclofen, dantrolene, baclofen, tizanidine, and dantrolene. Appropriate anti-inflammatory drugs include, but are not limited to, prednisone, nonsteroidal anti-inflammatory drugs (e.g., ibuprofen, naproxen, ketoprofen, and nimeslide), COX-2 inhibitors (e.g., rofecoxib, celecoxib, and etoricoxib), and immunoselective anti-inflammatory derivatives (e.g., submandibular peptide-T and its derivatives).

[0191] Appropriate antihistamines include H1 receptor antagonists (e.g., acribastine, azelastine, bilastine, brompheniramine, buclidine, bromodifenhydramine, carbinoxamine, cetirizine, chlorpromazine, cyclizine, chlorpheniramine, clemastine, cyproheptadine, desloratadine, dexbrompheniramine, dexchlorpheniramine, dimenhydrinate, dimethindene, diphenhydramine, doxylamine, ebastine, embramin, fexofenadine, hydroxy Examples include, but are not limited to, din, levocetirizine, loratadine, meclizine, mirtazapine, olopatadine, orphenadrine, phenindamine, pheniramine, phenyltroxamine, promethazine, pyriramine, quetiapine, rupatadine, triperenamine, and triprolidine), H2-receptor antagonists (e.g., cimetidine, famotidine, lafutidine, nizatidine, ranitidine, and roxatidine), tritocharin, catechin, cromoglycic acid, nedocromil, and p2-adrenergic agonists.

[0192] Appropriate antiinfective agents include amoebicides (e.g., nitazoxanide, paromomycin, metronidazole, tinidazole, chloroquine, miltefosine, amphotericin b, and iodoquinol), aminoglycosides (e.g., paromomycin, tobramycin, gentamicin, amikacin, kanamycin, and neomycin), anthelmintics (e.g., pyrantel, mebendazole, ivermectin, praziquantel, albendazole, thiabendazole, oxamnicine), antifungal agents (e.g., azole antifungals (e.g., itraconazole, fluconazole)), and (e.g., parconazole, ketoconazole, clotrimazole, miconazole, and voriconazole), echinocandins (e.g., caspofungin, anidurafungin, and micafungin), griseofulvin, terbinafine, flucytosine, and polyenes (e.g., nistatin and amphotericin b), antimalarial agents (e.g., pyrimethamine / sulfadoxine, artemata / lumefantrine, atovaquone / proguanil, quinine, hydroxychloroquine, mefloquine, chloroquine, doxycycline, pyrimethamine, and halofalamine), Antituberculosis agents (e.g., aminosalicylates (e.g., aminosalicylic acid), isoniazid / rifampin, isoniazid / pyrazinamide / rifampin, bedaquiline, isoniazid, ethambutol, rifampin, rifabutin, rifapentin, capreomycin, and cycloserine), antiviral agents (e.g., amantadine, rimantadine, abacavir / lamivudine, emtricitabine / tenofovir, cobicistat / elvitegravir / emtricitabine / tenofovir, efavirenz / emtricitabine / tenofovir, abacavir / Lamivudine / zidovudine, Lamivudine / zidovudine, Emtricitabine / tenofovir, Emtricitabine / lopinavir / ritonavir / tenofovir, Interferon α-2v / ribavirin, Pegylated interferon α-2b, Maraviroc, Raltegravir, Dolutegravir, Enfuvirtide, Foscarnet, Homivirsen, Oseltamivir, Zanamivir, Nevirapine, Efavirenz, Etravirine, Rilpivirine, Delavirdine, Nevirapine, Entecavir, Lamivudine, Adefovir, Sofosbuvir, Didanosine, Tenofovir, Abacavir, Zidovudine(Stabuzin, emtricitabine, zalcitabine, terbivudine, simeprevir, boceprevir, telaprevir, lopinavir / ritonavir, boceprevir, darunavir, ritonavir, tipranavir, atazanavir, nelfinavir, amprenavir, indinavir, saquinavir, ribavirin, valacyclovir, acyclovir, famciclovir, ganciclovir, and valganciclovir), carbapenems (e.g., doripenem, meropenem, ertapenem, and cilastatin / imipenam), cephalosporins (e.g., cefadroxil, cefradine) (e.g., cefazolin, cephalexin, cefepime, cefazolin, loracalbef, cefotetan, cefuroxime, cefprodil, loracalbef, cefoxitin, cefaclor, ceftibuten, ceftriaxone, cefotaxime, cefpodoxime, cefdinir, cefixime, cefditoren, ceftizoxime, and ceftazidime), glycopeptide antibiotics (e.g., vancomycin, dalbavancin, oritavancin, and teravancin), glycylcyclines (e.g., tigecycline), leprosy treatments (e.g., clofazimine and thalidomide) Lincomycin and its derivatives (e.g., clindamycin and lincomycin), macrolides and their derivatives (e.g., telithromycin, fidaxomicin, erythromycin, azithromycin, clarithromycin, zithromycin, and troleandomycin), linezolid, sulfamethoxazole / trimethoprim, rifaximin, chloramphenicol, fosfomycin, metronidazole, aztreonam, bacitracin, penicillin (amoxicillin, ampicillin, bacampicillin, carbenicillin, piperacillin, thi Calcillin, amoxicillin / clavulanate, ampicillin / sulbactam, piperacillin / tazobactam, clavulanate / ticalcillin, penicillin, procainepenicillin, oxacillin, dicloxacillin, and naphcillin), quinolones (e.g., lomefloxacin, norfloxacin, ofloxacin, gatifloxacin, moxifloxacin, ciprofloxacin, levofloxacin, gemifloxacin, moxifloxacin, cinoxacin, nalidixic acid, enoxacin, glepafloxacin, gatifloxacin, trovafloxacin,Examples include, but are not limited to, drugs such as bisphosphonates (and sparfloxacin), sulfonamides (e.g., sulfamethoxazole / trimethoprim, sulfasalazine, and sulfisoxazole), tetracyclines (e.g., doxycycline, demeclocycline, minocycline, doxycycline / salicylic acid, doxycycline / ω-3 polyunsaturated fatty acids, and tetracycline), and drugs for urinary tract infections (e.g., nitrofurantoin, methenamine, fosfomycin, cinoxacin, nalidixic acid, trimethoprim, and methylene blue).

[0193] Appropriate chemotherapy drugs include paclitaxel, brentuximab vedotin, doxorubicin, 5-FU (fluorouracil), everolimus, pemetrexed, melphalan, pamidronate, anastrozole, exemestane, nelarabine, ofatumumab, bevacizumab, bellinostat, tocitumomab, carmustine, bleomycin, bosutinib, busulfan, alemtuzumab, irinotecan, vandetanib, and bical. Tamide, Lomustine, Daunorubicin, Clofarabine, Cabozantinib, Dactinomycin, Ramucirumab, Cytarabine, Cytoxane, Cyclophosphamide, Decitabine, Dexamethasone, Docetaxel, Hydroxyurea, Dacarbazine, Leuprolide, Epirubicin, Oxaliplatin, Asparaginase, Estramustine, Cetuximab, Bismodegib, Asparaginase Erwinia chrysanthemis Chrysanthemi), amifostine, etoposide, flutamide, toremifene, fulvestrant, letrozole, degarelix, pralatrexate, methotrexate, phloxuridine, obinutuzumab, gemcitabine, afatinib, imatinib mesylate, carmustine, eribulin, trastuzumab, altretamine, topotecan, ponatinib, idarubicin, ifosfamide, ibrutinib, A Xitinib, interferon α-2a, gefitinib, romidepsin, ixabepyrone, ruxolitinib, cabazitaxel, adtrastuzumab ethansine, carfilzomib, chlorambucil, salglamostim, cladribine, mitotane, vincristine, procarbazine, megestrol, trametinib, mesna, strontium-89 chloride, mechloretamine, mitomycin, busulfan, gemtuzumabOzogamicin, vinorelbine, filgrastim, pegfilgrastim, sorafenib, nilutamide, pentostatin, tamoxifen, mitoxantrone, pegaspargase, denileukin difutitox, alitretinoin, carboplatin, pertuzumab, cisplatin, pomalidomide, prednisone, aldesleukin, mercaptopurine, zoledronic acid, lenalidomide, rituximab, octreotide, dasatinib, regorafenib, histrelin, sunitinib, siltuximab, omasetaxin, thioguanine, dabrafenib, erlotinib, bexarotene, temozolo Examples include, but are not limited to, mid, thiotepa, thalidomide, Bacillus calmette-guéran (BCG), temsirolimus, bendamustine hydrochloride, triptorelin, arsenic trioxide, lapatinib, barrubicin, panitumumab, vinblastine, bortezomib, tretinoin, azacitidine, pazopanib, teniposide, leucovorin, crizotinib, capecitabine, enzalutamide, ipilimumab, goserelin, vorinostat, idelalisib, ceritinib, abiraterone, epotilon, tafluposide, azathioprine, doxifluridine, vindesine, and all trans retinoic acid.

[0194] kit Any of the compounds and / or formulations described herein (e.g., P188 and P188-containing formulations) may exist as a combination kit. As used herein, the terms “combination kit” or “kit of components” refer to compounds, compositions, formulations, particles, cells, and any additional components used to package, sell, market, deliver, and / or administer a combination of elements or single elements (e.g., active ingredients) contained therein. Such additional components include, but are not limited to, packaging, syringes, blister packs, bottles, etc. If one or more of the compounds, compositions, formulations, particles, cells, or combinations thereof (e.g., drugs) contained in the kit are administered simultaneously, the combination kit may contain the active ingredient in a single formulation or separate formulations such as a pharmaceutical formulation (e.g., tablets, liquid preparations, dehydrated preparations, etc.). If the compounds, compositions, formulations, particles, and cells, or combinations thereof and / or kit components described herein are not administered simultaneously, the combination kit may contain each drug or other component in separate pharmaceutical formulations. Separate kit components may be contained in a single package within the kit or in separate packages.

[0195] In some embodiments, the combination kit also includes instructions printed on a tangible medium of expression or otherwise included. The instructions may provide information regarding the content of the compound and / or formulation, safety information regarding the content of the compound and formulation (e.g., pharmaceutical formulation), dosage, indications for use, and / or information regarding recommended treatment regimens for the compound and / or pharmaceutical formulation contained therein. In some embodiments, the instructions may provide instructions and protocols for administering the compound and / or formulation described herein to subjects requiring it. In some embodiments, the instructions may provide one or more embodiments of methods for administering P188 and / or its pharmaceutical formulation, such as any of the methods described in more detail elsewhere in this specification.

[0196] Methods to prevent non-irradiated cell damage during radiation exposure Some exemplary embodiments of this specification describe methods for preventing non-injured cellular damage during and / or after injury. As previously stated, injury can be any injury, e.g., energetic injury (e.g., radiation), chemical injury, biological injury (e.g., virus or microorganism), physiological injury (e.g., ischemic event such as myocardial infarction or stroke), and / or mechanical injury (e.g., heat, cold, pressure, melting, etc.). Exposure to injury may be intentional, such as for treatment, or accidental, such as unintentional exposure to injury in the environment. Injury may be unplanned or unexpected, as in the case of physiological injury.

[0197] Generally, the method may include administering one or more doses of P188 or its pharmaceutical formulations before, during, and / or after injury. One or more doses of P188 or its pharmaceutical formulations may be administered 0 to 48 hours before and / or 0 to 48 hours after injury. In some embodiments, it is preferable to administer one or more doses of P188 before exposure to injury. In some embodiments, it is understood that post-injury administration of P188 may not provide as much benefit as pre-injury administration of P188, but nevertheless, benefits may be achieved. It will also be understood that administering one or more doses of P188 after injury may be advantageous compared to not administering any dose at all, especially when exposure to injury is unpredictable or unplanned (e.g., in the case of an ischemic event or accidental radiation exposure). In some embodiments, the one or more doses of P188 given after injury are given as close as possible to the time of injury. For example, during an ischemic event, this may be before catheter placement and tpa.

[0198] In some exemplary embodiments, the injury is radiation, and therefore, methods for preventing non-irradiated cell damage during radiation, such as during radiotherapy, are described herein. Figures 8A–8C illustrate embodiments of the methods described below in further detail.

[0199] In some embodiments, the method may include administering a first amount of P188 or a pharmaceutical formulation thereof to one or more areas of the subject before exposing them to injury; and exposing one or more areas of the subject to injury after the blood concentration of P188 has reached 1 and about 5 mg / mL (e.g., about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or about 5 mg / mL). As shown in Figure 8A, the first dose may be a single bolus dose, so that at a specific time after administration, the blood concentration of P188 reaches a concentration of 1 to about 5 mg / mL (e.g., about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or about 5 mg / mL), at which point one or more areas of the target may be exposed to injury. In some embodiments, the areas exposed to injury include one or more cancerous cells. In some embodiments, as shown in Figure 8A, the subject receives a first amount of P188 or its pharmaceutical formulation and does not receive any further amounts of P188 or its pharmaceutical formulation for at least about 14 to 24 hours.

[0200] In some embodiments, the first dose, when given as a single bolus dose, is such that the blood concentration of P188 reaches a concentration of 1 to about 5 mg / mL (e.g., about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or about 5 mg / mL) within about 1 to about 48 hours. In some embodiments, when the first dose is given as a single bolus dose, the blood concentration of P188 is 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8 0.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11.75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.7 5, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 20.25, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, 24, 24.25, 24.5, 24.75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26.5, 26.75, 27, 27 0.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29.25, 29.5, 29.75, 30, 30.25, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32.75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75, 36, 36.25, 36.5, 36.75, 37, 37.25, 37.5, 37.75, 38, 38.25, 38.5, 38.75, 39, 39.25, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42.5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47 The concentration reaches approximately 1 to 5 mg / mL (for example, approximately 0, 0.25, 0.5, 0.75, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or approximately 5 mg / mL) after 0.75 or approximately 48 hours. Therefore, in some embodiments, one or more areas of a subject can be exposed to ionizing radiation approximately 0 hours (i.e., simultaneously with the injury) to 48 hours after administration of a first amount of P188 or its pharmaceutical formulation. In some embodiments, one or more regions of the target area are approximately 0, 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11 11.25, 11.5, 11.75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.75, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 20.25, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, 24, 24.25, 24.5, 24.75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26.5, 26.75, 27, 27.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29 0.25, 29.5, 29.75, 30, 30.25, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32.75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75 36, 36.25, 36.5, 36.75, 37, 37.25, 37.5, 37.75, 38, 38.25, 38.5, 38.75, 39, 39.25, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42. Exposure to ionizing radiation may occur at 5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47.75, or approximately 48 hours later.

[0201] In some embodiments, the blood concentration of P188 or its pharmaceutical formulation can be measured at one or more time points after the administration of a first dose to determine when one or more areas of the subject should be exposed to radiation. In some embodiments, the time for exposing one or more areas of the subject to radiation may be calculated based, for example, the amount of P188 administered, the half-life of P188 (which is about 4.5 hours), and the characteristics of the subject (e.g., age, weight, medical condition, and others as understood by the physician).

[0202] As shown in Figure 8B, multiple bolus doses of P188 or its pharmaceutical formulation can be administered. The total number of additional doses is designated as "n" elsewhere in this specification and in Figure 8B. Any specific dose after the first amount given as a bolus dose is designated as "n" th This can be called the dosage of '. For example, dosage 5 out of a total of 8 units. thIt can be called the dose. In some situations, n th The dose may refer to the last dose administered. In some embodiments, the number of doses that may be administered after the first dose may be 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In some embodiments, when given as a single dose, any dose is a P188 blood concentration of 1 to about 5 mg / mL (e.g., about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or about 5 mg / mL). The goal is to achieve a cumulative P188 blood concentration of 1 to approximately 5 mg / mL (e.g., approximately 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or approximately 5 mg / mL), although this may be less than what would be achieved. Similar to those discussed with respect to the embodiments illustrated in Figure 8A, the amount required for each bolus dose administered can be determined by measuring the P188 concentration in the bloodstream, or it can be calculated based, for example, the amount of P188 administered, the half-life of P188 (which is about 4.5 hours), and the characteristics of the subject (e.g., age, weight, medical condition, and others recognized by the physician).

[0203] As illustrated and discussed with respect to Figure 8A, the total amount of P188 administered over all bolus doses, the blood concentration of P188, is 1, n th, or within approximately 1 to 48 hours of the last dose, for example, approximately 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11.75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.75, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 20.2 5, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, 24, 24.25, 24.5, 24.75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26. 5, 26.75, 27, 27.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29.25, 29.5, 29.75, 30, 30.25, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32. 75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75, 36, 36.25, 36.5, 36.75, 37, 37.25, 37.5, 37.75, 38, 38.25, 38.5, 38.75, 39 , 39.25, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42.5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47.75, or within approximately 48 hours, 1 to approximately 5 mg / mL (for example, approximately 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.The concentration reaches 2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or approximately 5 mg / mL). Therefore, in some embodiments, one or more areas of the target reach a first amount, n. th The amount of P188 or the last dose of its pharmaceutical formulation may be used to expose the area to injury approximately 0 hours (i.e., simultaneously with injury exposure) to 48 hours after administration. In some embodiments, one or more areas of the area are exposed to the first dose, n thThe dosages of P188 or the last dose of the pharmaceutical formulation are approximately 0, 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11.75, 12, 12.25, 12.5 ,12.75,13,13.25,13.5,13.75,14,14.25,14.5,14.75,15,15.25,15.5,15.75,16,16.25,16.5,16.75,17,17.25,17.5,17.75,18,18.25,18.5,18.75,19,19.25,19.5,19.75,20,20.25,20.5,20.75,21,21.25,21.5,21.75,22,22.25,22.5,22.75,23,23.25,23.5,23.75,24,24.25,24.5,2 4.75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26.5, 26.75, 27, 27.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29.25, 29.5, 29.75, 30, 30.25, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32.75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75, 36, 36.25, 36.5, 36. You may be exposed to injury at 75, 37, 37.25, 37.5, 37.75, 38, 38.25, 38.5, 38.75, 39, 39.25, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42.5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47.75, or approximately 48 hours later.

[0204] In some embodiments, the subject does not receive further P188 for at least 14 to 24 hours after the last bolus dose of P188 or the pharmaceutical formulation has been administered.

[0205] As illustrated in Figure 8C, P188 or its pharmaceutical formulation can be continuously injected into a subject over a period of time before one or more areas of the subject are exposed to injury.In some embodiments, a certain period is approximately 0 to 48 hours, for example, approximately 0, 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11.75, 12, 12.25, 1 2.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.75, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 20.25, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, 24, 24.25, 24 0.5, 24.75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26.5, 26.75, 27, 27.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29.25, 29.5, 29.75, 30, 30.25, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32.75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75, 36, 36.25, 36. 5, 36.75, 37, 37.25, 37.5, 37.75, 38, 38.25, 38.5, 38.75, 39, 39.25, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42.5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47.75, or approximately 48 hours.In some embodiments, P188 or its pharmaceutical formulations may be continuously infused into a subject until the blood concentration of P188 reaches a concentration of approximately 1 to approximately 5 mg / mL (e.g., approximately 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or approximately 5 mg / mL).In some embodiments, P188 blood concentrations are within approximately 1 to 48 hours, for example, approximately 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11.75, 12, 12.25, 12.5, 12. 75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.75, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 20.25, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, 24, 24.25, 24.5, 24. 75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26.5, 26.75, 27, 27.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29.25, 29.5, 29.75, 30, 30.25, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32.75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75, 36, 36.25, 36.5, 36. 75, 37, 37.25, 37.5, 37.75, 38, 38.25, 38.5, 38.75, 39, 39.25, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42.5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47.75, or in about 48 hours, this amount is reached.

[0206] In some embodiments, a method for preventing damage to non-injured cells in a subject during and / or after injury exposure may include administering a certain amount of poloxamer 188 (P188) or a pharmaceutical formulation thereof to one or more areas of the subject before exposure of those areas to injury, the amount being in the range of about 10 to about 150 mg / kg; and exposing one or more areas of the subject to ionizing radiation (or other injury) within 0 to 48 hours after administration of a certain amount of P188 or a pharmaceutical formulation thereof. In some embodiments, the amount may be in the range of about 100 to 150 mg / kg. In some embodiments, the amount may be in the range of about 20 to 40 mg / kg. In some embodiments, a first dose (e.g., loading dose) may be in the range of about 100 to about 150 mg / kg. In some embodiments, a second dose may be delivered once or more times for maintenance, and may be in the range of about 20 to about 40 mg / kg. In some embodiments, the maintenance dose may be administered once or more times per hour.In some embodiments, exposure to injury in one or more areas of the target area is approximately 0, 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11. 75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.75, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 20.25, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, 24, 2 4.25, 24.5, 24.75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26.5, 26.75, 27, 27.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29.25, 29.5, 29.75, 30, 30.2 5, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32.75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75, 36, 36.25, 36 0.5, 36.75, 37, 37.25, 37.5, 37.75, 38, 38.25, 38.5, 38.75, 39, 39.25, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42.5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47.75, or may be performed within approximately 48 hours.

[0207] In some embodiments, injured cells may be diseased cells, abnormal cells, or otherwise unhealthy cells. In some embodiments, injured cells may be cancerous. In some embodiments, injured cells may be non-cancerous tumors or non-tumor cells. Non-injured cells may be normal cells or otherwise healthy cells. In some embodiments, non-injured cells are soft tissue cells. In some embodiments, non-injured cells are vascular cells. In some embodiments, non-injured cells are endothelial cells.

[0208] Methods for treating diseases or conditions are also described herein. In some embodiments, the disease or condition may be caused by injury to cells or tissues. In some embodiments, the disease or condition may be a disease or condition that causes the production of free radicals or oxidation in / by affected cells and / or stimulates an inflammatory response or cascade. In some embodiments, treatment of the disease or condition may cause injury to cells or tissues. In some embodiments, the disease or condition may be a virus. In some embodiments, the disease or condition may be acute respiratory distress syndrome (ARDS). In some embodiments, the disease or condition may be a cardiac condition. In some embodiments, the disease or condition may be a neurological disease or condition. In some embodiments, the disease or condition may be acute radiation sickness (ARS) (see, for example, Williams and McBride, WH2011. Int Radiat Biol 87(8):851-868). In some embodiments, the disease or disorder may be radiotoxicity. In some embodiments, the disease or disorder may be an injury such as radiation-induced pneumonia. In some embodiments, the disease or disorder is an ischemic event (e.g., stroke or myocardial infarction).

[0209] In some embodiments, a method for treating a disease or condition may include administering a first dose of poloxamer 188 (P188) or a pharmaceutical formulation thereof to one or more areas of a subject before exposing those areas to injury; and then exposing one or more areas of a subject to injury after the blood concentration of P188 has reached approximately 1 to approximately 5 mg / mL (e.g., approximately 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or approximately 5 mg / mL). In some embodiments, the method involves administering a first dose of poloxamer 188 (P188) or a pharmaceutical formulation thereof to a subject before the disease or condition is exposed to injury in one or more areas of the subject; and administering doses of 0, 0.25, 0.5, 0.75, 1, 1, 1, 48 hours after the administration of the first dose of P188 or the pharmaceutical formulation, for example, from the time of injury. 25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11 0.5, 11.75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.75, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 2 0, 20.25, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, 24, 24.25, 24.5, 24.75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26.5, 26.75, 27, 27.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29.25, 29.5, 29.75, 30, 30.25, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32.75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75, 36, 36.25, 36.5, 36.75, 37, 37.25, 37.5, 37.75, 38, 38.25, 38.5, 38.75, 39, 39.2 This may include exposing one or more areas of the subject to injury at 5, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42.5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47.75, or approximately 48 hours later. In some embodiments, the method may include administering multiple doses of P188 or its pharmaceutical formulation, similar to those described above with respect to Figure 8B. In some embodiments, the method may include administering P188 or its pharmaceutical formulation in a manner similar to that described above with respect to Figure 8C.

[0210] Methods for treating non-cancerous tumors or conditions are also described herein. In some embodiments, a method for treating a non-cancerous tumor or condition involves administering a first dose of poloxamer 188 (P188) or a pharmaceutical formulation thereof to one or more areas of the subject before exposure of the subject to ionizing radiation; and the blood concentration of P188 being approximately 1 to approximately 5 mg / mL (e.g., approximately 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or about 5 mg / mL) can be included in exposing one or more areas of the subject to ionizing radiation. In some embodiments, a method for treating a non-cancerous tumor or condition may include administering a first dose of poloxamer 188 (P188) or a pharmaceutical formulation thereof to the subject before exposure of one or more areas of the subject to ionizing radiation; and administering, for example, about 0, 0.25, 0 0 0 0 48 hours after administration of the first dose of P188 or the pharmaceutical formulation. 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11.75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.75, 18, 18.25, 1 8.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 20.25, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, 24, 24.25, 24.5, 24.75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26.5, 26.75, 27, 27.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29.25, 29.5, 29.75, 30, 30.25, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32.75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75, 36, 36.25, 36.5, 36.75, 37, 37.25, 37.5, 37.75, 38, 38.25 This may include exposing one or more areas of the subject to ionizing radiation at 38.5, 38.75, 39, 39.25, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42.5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47.75, or approximately 48 hours later. In some embodiments, the method may include administering multiple doses of P188 or its pharmaceutical formulation, similar to those described above with respect to Figure 8B. In some embodiments, the method may include administering P188 or its pharmaceutical formulation in a manner similar to that described above with respect to Figure 8C.

[0211] In some embodiments, non-cancerous tumors (i.e., benign tumors) or other non-cancerous conditions may be acoustic neuromas, arteriovenous malformations, thyroid disorders, trigeminal neuralgia, meningiomas, inflammatory / proliferative disorders (e.g., Dupuytren's disease, ectopic ossification, keloid scars, pigmented villonodular synovitis), other benign tumors, and other conditions (e.g., McKeown et al., (Br J.Radiol.2015 88(1056), in particular Table 1; see also Seegenschmiedt et al. Br J Radiol.2015:88(1051)).

[0212] Methods for treating cancer are also described in this specification. In some embodiments, a method for treating cancer may include administering a first dose of poloxamer 188 (P188) or a pharmaceutical formulation thereof to one or more areas of the area before exposing them to ionizing radiation; and exposing one or more areas of the area to ionizing radiation after the blood concentration of P188 has reached 1 to about 5 mg / mL (e.g., about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or about 5 mg / mL). In some embodiments, a method of treating cancer involves administering a first dose of poloxamer 188 (P188) or a pharmaceutical formulation thereof to one or more areas of the subject before exposure to ionizing radiation; and administering 0 to 48 hours after the administration of the first dose of P188 or the pharmaceutical formulation, for example, about 0 (i.e., delivery at time of injury) 0.25, 0. 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11.75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.75, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 20.25, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, 24, 24.25, 24.5, 24.75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26.5, 26.75, 27, 27.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29.25, 29.5, 29.75, 30, 30.25, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32.75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75, 36, 36.25, 36.5, 36.75, 37, 37.25, 37.5, 37.75, 38, 38.25, 38.5, 3 This may include exposing one or more areas of the subject to ionizing radiation at 8.75, 39, 39.25, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42.5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47.75, or approximately 48 hours later. In some embodiments, the method may include administering multiple doses of P188 or its pharmaceutical formulation, similar to those described above with respect to Figure 8B. In some embodiments, the method may include administering P188 or its pharmaceutical formulation in a manner similar to that described above with respect to Figure 8C.

[0213] In some embodiments, cancer is a solid tumor cancer. In some embodiments, cancer is a non-solid tumor. In some embodiments, cancer is a spinal cord tumor, vertebral tumor, leukemia, head and neck cancer, Ewing's sarcoma, soft tissue sarcoma, prostate cancer, esophageal cancer, colorectal cancer, paranasal sinus cancer, pancreatic cancer, chordoma, osteosarcoma, chondrosarcoma, breast cancer, meningioma, brain tumor, bone cancer, lung cancer, lymphoma (including, but not limited to, non-Hodgkin lymphoma), liver cancer, or a combination thereof.

[0214] In some embodiments, the amount of ionizing radiation to which one or more areas of the target are exposed is less than or equal to about 100 Gy, but greater than zero. In some embodiments, the amount of ionizing radiation to which one or more areas of the target are exposed is about 1 to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 The range is 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 Gy. In some embodiments, the amount of ionizing radiation to which one or more areas of a target are exposed is approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 4 7, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 Gy.

[0215] In some embodiments, the amount of ionizing radiation to which one or more areas of the target are exposed is less than or equal to about 80 Gy, but greater than zero. In some embodiments, the amount of ionizing radiation to which one or more areas of the target are exposed is about 1 to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 3 The range is 7, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, and 80 Gy. In some embodiments, the amount of ionizing radiation to which one or more areas of a target are exposed is approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 3 7, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80 Gy.

[0216] In some embodiments, the amount of ionizing radiation to which one or more areas of the target are exposed is 50 Gy or less, but greater than zero. In some embodiments, the amount of ionizing radiation to which one or more areas of the target are exposed is in the range of about 3 to about 50 Gy, for example, about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or about 50 Gy. In some embodiments, the amount of ionizing radiation to which one or more areas of the target are exposed may be 50 Gy or more. In some embodiments, the amount of ionizing radiation to which one or more areas of the subject are exposed may be about 50 to about 100 Gy, for example, about 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or about 100 Gy. In some embodiments, exposure to ionizing radiation is accidental. In some embodiments, the subject has developed or is at risk of developing acute radiation syndrome. In some embodiments, the subject may be exposed to >0.05 Gy (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.0, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or about 50 Gy).

[0217] Methods for preventing injury-induced pneumonia in subjects are also described herein. In some embodiments, the methods include a) administering a certain amount of P188 or a formulation thereof to a subject before exposure to injury; b) administering a certain amount of P188 or a formulation thereof to a subject immediately after exposure to injury; c) administering a certain amount of P188 or a formulation thereof to a subject during injury; or d) a combination thereof. In some embodiments, certain amounts are effective in increasing the blood concentration of P188 in a subject to 1 mg / mL to 5 mg / mL (e.g., about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or about 5 mg / mL). In some embodiments, certain amounts are effective in coating one or more non-harmful cells. In some embodiments, certain amounts are effective in reducing or preventing oxidative damage to one or more non-harmful cells. In some embodiments, a certain amount is effective in reducing or preventing inflammatory damage to one or more non-injured cells.In some embodiments, administration is given 0 to 48 hours before exposure to injury, for example, about 0 (i.e., administration at the time of injury) 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 1 1.75, 12, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.7 5, 18, 18.25, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 20.25, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, 2 4, 24.25, 24.5, 24.75, 25, 25.25, 25.5, 25.75, 26, 26.25, 26.5, 26.75, 27, 27.25, 27.5, 27.75, 28, 28.25, 28.5, 28.75, 29, 29.25, 29.5, 29.75, 30, 3 0.25, 30.5, 30.75, 31, 31.25, 31.5, 31.75, 32, 32.25, 32.5, 32.75, 33, 33.25, 33.5, 33.75, 34, 34.25, 34.5, 34.75, 35, 35.25, 35.5, 35.75, 36, 36.2 It takes place at 5, 36.5, 36.75, 37, 37.25, 37.5, 37.75, 38, 38.25, 38.5, 38.75, 39, 39.25, 39.5, 39.75, 40, 40.25, 40.5, 40.75, 41, 41.25, 41.5, 41.75, 42, 42.25, 42.5, 42.75, 43, 43.25, 43.5, 43.75, 44, 44.25, 44.5, 44.75, 45, 45.25, 45.5, 45.75, 46, 46.25, 46.5, 46.75, 47, 47.25, 47.5, 47.75, or approximately 48 hours prior.In some embodiments, administration is from 0 to 24 hours after exposure to injury (i.e., administration at the time of injury), for example, approximately 0, 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, 9.75, 10, 10.25, 10.5, 10.75, 11, 11.25, 11.5, 11.75, 1 2, 12.25, 12.5, 12.75, 13, 13.25, 13.5, 13.75, 14, 14.25, 14.5, 14.75, 15, 15.25, 15.5, 15.75, 16, 16.25, 16.5, 16.75, 17, 17.25, 17.5, 17.75, 18, 18.2 The process takes place over 5, 18.5, 18.75, 19, 19.25, 19.5, 19.75, 20, 20.25, 20.5, 20.75, 21, 21.25, 21.5, 21.75, 22, 22.25, 22.5, 22.75, 23, 23.25, 23.5, 23.75, or approximately 24 hours. In some embodiments, the injury is mechanical, chemical, biological, energetic, or a combination thereof. In some embodiments, the injury is ionizing radiation.

[0218] In some embodiments, the amount of ionizing radiation is less than or equal to about 100 Gy, but greater than zero. In some embodiments, the amount of ionizing radiation to which one or more areas of the target are exposed is about 1 to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 The range is 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 Gy. In some embodiments, the amount of ionization is approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 , 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 Gy.

[0219] In some embodiments, the amount of ionizing radiation is less than or equal to approximately 80 Gy, but greater than zero. In some embodiments, the amount of ionizing radiation is in the range of approximately 1 to approximately 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80 Gy. In some embodiments, the amount of ionizing radiation is approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80 Gy.

[0220] In some embodiments, the amount of ionizing radiation is 50 Gy or less. In some embodiments, the amount of ionizing radiation may be in the range of about 3 to about 50 Gy, for example, about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or about 50 Gy. In some embodiments, the amount of ionizing radiation may be 50 Gy or more. In some embodiments, the amount of ionizing radiation may be about 50 to about 100 Gy, for example, about 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or about 100 Gy. In some embodiments, exposure to ionizing radiation is accidental. In some embodiments, the subject has developed or is at risk of developing acute radiation syndrome. In some embodiments, the subject may be exposed to >0.05 Gy (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or about 50 Gy).

[0221] In some embodiments, in addition to pretreatment with the P188 formulation as described above, the method may also include administering an amount of the P188 formulation described herein immediately after injury (e.g., within 1 to 24 hours, and / or when the injury is still in the initial phase and proximal / distant cell damage is still mediated by oxidation).

[0222] In some embodiments, particularly when exposure to injury cannot be predicted and therefore pretreatment with P188 or its pharmaceutical formulation is unavailable, one or more doses of P188 or its pharmaceutical formulation are administered once or twice between 0 and 48 hours post-injury, for example, at approximately 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or approximately 48 hours post-injury. Exemplary unpredictable injury exposures for which simultaneous or post-injury administration of P188 is the only option include, for example, physiological events (e.g., ischemic events) and accidental radiation exposure. Other matters will be understood in light of the disclosures herein.

[0223] In some embodiments, the method may include administering a co-therapy, such as an adjuvant activator, which may be administered as a standalone pharmaceutical formulation or included in the poloxamer formulation. When co-therapy, adjuvants, and / or multiple pharmaceutical formulations are delivered to a subject and / or cells, the different therapies or formulations may be administered sequentially or simultaneously. Sequential administration is administration where a considerable amount of time occurs between administrations, such as approximately 15, 20, 30, 45, 60 minutes or more. The time between administrations in sequential administration may be on the order of hours, days, months, or years, depending on the activators present in each administration. Co-administration refers to administering two or more formulations simultaneously or substantially simultaneously (e.g., within seconds or minutes apart), with the intention of administering the formulations together at the same time. In some embodiments, adjuvants, co-therapy, etc., may be administered before, after, or both before and after the poloxamer formulation. In some embodiments, the method described herein is a co-therapy for treating injury, such as biological injury. In some embodiments, the method described herein is a combination therapy in which the primary treatment causes injury to cells, such as affected cells.

[0224] Further embodiments are illustrated in the following examples, which are provided for illustrative purposes only and are not intended to limit the scope of the invention. [Examples]

[0225] While embodiments of this disclosure have been described, the following examples generally describe several additional embodiments of this disclosure. Embodiments of this disclosure are described in relation to the following examples and corresponding text and figures, but are not intended to limit the embodiments of this disclosure to this description. Rather, they are intended to cover all substitutes, modifications, and equivalents that fall within the spirit and scope of the embodiments of this disclosure. The following examples are provided to give a complete disclosure and description of how to carry out the methods disclosed and claimed herein, and how to use the probe. Efforts have been made to ensure accuracy with respect to numbers (e.g., quantity, temperature, etc.), but some error and deviation should be taken into consideration. Unless otherwise specified, parts are parts by weight, temperatures are °C, and pressures are atmospheric pressure or near atmospheric pressure. Standard temperature and pressure are defined as 20 °C and 1 atm.

[0226] Example 1 - Hiroloxamer protects cells from hydrogen peroxide. During radiation therapy for cancer treatment, the vascular system is inevitably exposed to radiation. Ionizing radiation causes cell death, either directly by damaging cellular DNA or indirectly by producing harmful byproducts such as free radicals and reactive oxygen species. The vascular system responds to ionizing radiation in two phases. The acute / initial phase occurs within 24 hours, when directly damaged cells undergo cell death, which impairs the barrier integrity of the vascular system. Cells indirectly exposed to byproducts send out damage signals and induce a pro-inflammatory response. If these complications do not resolve, later effects (including capillary collapse and scar formation) begin to appear.

[0227] As can be seen at least by Figure 1, Hiroloxamer was observed to provide protection to human adipose microvascular endothelial cells when exposed to hydrogen peroxide, preventing damage to these cells. Briefly, human adipose microvascular endothelial cells (HAMECs) were stained with MitoTracker Deep Red FM to label functional mitochondria and seeded into 12-well plates. The cells were allowed to adhere and grow to confluence. Once confluenced, appropriate wells were treated with Hiroloxamer for 3 hours. After 3 hours of incubation, 1 mM hydrogen peroxide was added to each well overnight. The ability of Hiroloxamer to provide protection to endothelial cells exposed to hydrogen peroxide was demonstrated by the presence of a complex mitochondrial network (Figure 1, grayscale) observed in the Hiroloxamer-treated group, which was not observed in the untreated group.

[0228] Example 2 - Hiroloxamer does not interfere with the radiation effect on cancer cells. In the context of radiotherapy, it is important that Hiroloxamer treatment does not interfere with the ability of radiation to effectively kill cancer cells. Here, it can be shown that Hiroloxamer treatment before radiotherapy does not interfere with the effect of radiation on cancer cells (Figures 2A-2B and 36A-36B). Using two cancer cell lines, H460 and A549, we verified that Hiroloxamer treatment before radiotherapy does not interfere with the ability of RT to effectively kill cancer cells. Briefly, cells were cultured and exposed to specified concentrations of Hiroloxamer for 3 hours before exposure to 0, 2, or 6 Gy of ionizing radiation. Survival rates were then determined for each group. There was no significant difference between the control group and the treated group, nor was there a significant difference in Hiroloxamer concentration.

[0229] Example 3 - Effects of Hiroloxamer on healthy microvascular endothelial cells. Following the results shown in Example 2, which demonstrated that Hiroloxamer administration does not affect the effectiveness of radiotherapy for killing cancer cells, its effect on healthy human adipose microvascular endothelial cells (HAMECs) was investigated. Cells were seeded in the wells of 12-well plates and incubated for 2 days prior to irradiation. To investigate the effect of Hiroloxamer as a prophylactic and palliative agent, 0.2 mM Hiroloxamer in Endothelial Growth Medium-2 (EGM2, Lonza) was administered at various time points: approximately 3 hours before irradiation (Rx pretreatment), approximately 30 minutes after irradiation (Rx@0.5), and approximately 24 hours after irradiation (Rx@24). Each 12-well plate was placed in an X-Rad320 and directly exposed to X-ray irradiation of 0.5, 1, 2, 4, 10, 20, or 50 Gy. After irradiation, the Rx@0.5 group was supplemented with a dose-appropriate amount of the drug and incubated overnight. The following day, all media were replaced and refreshed with EGM-2. At this point, Rx@24Hiroloxamer was administered, and the Hiroloxamer was removed the following day. Hiroloxamer was administered to each group for 24 hours, then removed and replaced with fresh EGM-2. Live cell imaging was performed with LionHeartFX every 24 hours throughout the study. Plates were restored to confluence and fixed in 2% PFA for analysis. Wells were stained for the nuclei with Phalloidin 633 (F-Actin) and Hoescht.

[0230] Similar to how endothelial cells organize within the vascular system, they form dense cell-cell adhesions in a dish, creating a compact monolayer across the entire dish. The cells possess an internal cytoskeleton composed primarily of F-actin, which regulates cell shape, provides strength, and is highly dynamic in response to cellular needs. In a confluent monolayer, the internal cytoskeletal structure of the cells is unidirectional and anisotropically homogeneous. In these monolayers and in the vascular system, endothelial cells are directly linked via adhesion complexes (adhesion junctions) that directly connect to the internal cytoskeleton. These complexes provide barriers that regulate vascular permeability. The distinct green outlines of each individual cell in the panel controls of Figures 3A–3P are cortical actin borders that determine cell shape. These are associated with cell-cell and cell-matrix adhesion complexes that keep cells firmly adhered to each other and to the matrix (e.g., for maintaining endothelial barrier integrity that regulates vascular permeability). Ionizing radiation is known to damage the endothelium and disrupt its barrier function through the destruction of cell-cell adhesions and the actin cytoskeleton. Human adipose microvascular endothelial cells directly exposed to 0.5 Gy of X-ray irradiation recovered to a confluent monolayer with typical F-actin expression, regardless of drug treatment (Figure 3A-3B, panel 0.5 Gy). It has been reported that cells recover to a confluent monolayer at this dose.

[0231] Moderate doses of irradiation (1–4 Gy) induce actin rearrangement from its cortical distribution to tension fibers extending throughout the cell body. This remodeling of the cell's cytoskeleton leads to cell contraction and loss of intercellular junctions, resulting in increased vascular permeability in vivo. This effect was observed in cells exposed to irradiation without Hiroloxamer pretreatment (Figure 3A–3B, panels 1, 2, 4 Gy). However, in the Hiroloxamer pretreatment group, cells re-established cortical actin margins and formed dense cell-cell junctions with no apparent intercellular gaps. Higher doses of ionizing radiation (approximately 10–50 Gy) result in increased cell surface area in the group, likely due to loss of intercellular adhesion and secondary damage from indirect radiation effects (Figure 3A–3B, panels 10, 20, 50 Gy). The 10 Gy Hiroloxamer group had more cells that appeared to have no increased cell surface area, were activated and mobile, and potentially attempted to re-establish cell-cell adhesion lost during exposure (Figure 3A-3B, panel 10 Gy). At higher doses, 20 and 50 Gy, there was a clear increase in cell surface area in all groups, but Hiroloxamer pretreatment appeared to prevent bleving and cell membrane bursting, and cells appeared rounder and more intact compared to the untreated group (Figure 3A-3B, panels 20, 50 Gy).

[0232] Example 4 - Protection of non-irradiated cells adjacent to irradiated cells This example may demonstrate that Hiroloxamer can protect human adipose microvascular endothelial cells directly adjacent to areas exposed to ionizing radiation. Endothelial cells were stained with MitoTracker Deep Red to label mitochondria for live-cell imaging. HAMEC cells were seeded in 60 mm petri dishes and grown to confluence. Three hours before irradiation, Hiroloxamer was added to appropriate dishes. Each petri dish was placed in an irradiator with a lead shield positioned at the top with a 6 mm hole for radiation exposure, exposing cells in this area (10% of the total plate area) to 10 Gy of radiation. After irradiation, the plates were returned to the incubator overnight. The following day, the medium was changed with normal growth medium, and all plates were fixed 48 hours after irradiation. Mitochondrial potentials were evaluated in different areas after irradiation: within the irradiated / exposed (6 mm) area, immediately adjacent to the exposed area, and far from the exposed area. The intensity of MitoTracker Deep Red (MTDR) changes with mitochondrial potential; viable cells appear as highly fluorescent clusters, while apoptotic cells exhibit lower fluorescence. The intensity and structure of mitochondria indicated by MitoTracker Deep Red dye were compared with corresponding phase-contrast images to check the integrity of the cell membrane. Cells with low MTDR signals and damaged cell membranes were considered dead.

[0233] Within the irradiated area, i.e., the area where the irradiation could pass through the 6 mm shield, there was no difference in cell viability between the different groups (Figures 4A-4E; n=16; p<0.974). This was an expected result, as the Hiroloxamer treatment did not affect the cancer cells that were directly exposed.

[0234] Survival rates were also evaluated in areas directly adjacent to the irradiation area but outside the 6 mm exposure zone. In areas close to the irradiation area, there was a significant increase in the number of MitoTracker Deep Red-positive cells with intact cell membranes, as confirmed by corresponding phase-contrast imaging, in the Hiroloxamer-treated group compared to the untreated control group (n=9; p<0.002; Figures 5A-5E).

[0235] Finally, we compared the regions of the dish furthest from the irradiation area (Figures 6A-6E). Compared to the control group, the Hiroloxamer-treated group showed a significant increase in MitoTracker Deep Red-positive cells (n=4, p<0.030; Figures 5A-5E). The sample size in this group was smaller due to the limited size of the dishes and the maintenance of a distance of at least 30 mm from the irradiation area.

[0236] Example 5 - Effect of pre-irradiation with Hiroloxamer on 3D tissue constructs. The effect of administering Hiroloxamer before radiation exposure to three-dimensional tissue engineering constructs was investigated. Previous reports on these constructs exist (Pattanaik, S., et.al. 2018). Scaffold-free prevascularized endothelial-fibroblast constructs (SPECs) are formed by seeding normal human dermal fibroblasts and human adipose microvascular endothelial cells (4:1) into non-adhesive agarose molds and culturing them in normal medium (FGM-2:EGM-2, 2:1) for 4 days. SPECs were seeded, and Hiroloxamer was administered on day 3, 3 hours before radiation exposure. The constructs were exposed to 10 Gy of X-rays in an X-Rad320 irradiator set to 320.0 kV and 12.50 mA. The constructs were returned to an incubator overnight and fixed with 4% PFA the following day. Next, the constructs were stained with Phalloidin 633, Hoescht, and CD31 for analysis of the actin cytoskeleton and endothelial network within the constructs. All constructs were imaged using a Leica SP5 confocal microscope with the same channel configuration and similar stack size.

[0237] Figures 7I–7L show Rx pretreatment (88.031 μm with a 4 μm step size) exposed to 10 Gy. In the control SPEC (no Rx, no XRT), a lace-like endothelial network (red) is present throughout the construct, embedded in a rich and consistent F-actin network (green) (Figures 7A–7D). The formed endothelial network resembles the primitive capillary-like network in vivo (Pattanaik). When the SPEC was exposed to 10 Gy of X-ray irradiation, there was clear actin depolymerization (indicated by a decrease in F-actin expression), and the endothelial network expanded and became disordered (Figures 7E–7H). The changes in the vascular component of the construct are similar to those seen in the vascular system in vivo in response to ionizing radiation, with loss of cell-cell adhesion and increased vascular permeability. The increased size of the vascular network in the irradiated SPEC may be due to the loss of cytoskeletal components, which help maintain the structure of the construct while losing adhesion between endothelial cells. However, when Hiroloxamer was added before irradiation, a lace-like endothelial network extending throughout the entire construct, embedded in a high-density F-actin network, remained present, similar to what was observed in the control unirradiated SPEC. These results suggest that Hiroloxamer helps preserve the actin cytoskeleton and also helps maintain cell-cell junctions after exposure to ionizing radiation.

[0238] Figures 9A-9C show fluorescence images that demonstrate the effect of Hiorloxamer pretreatment on F-actin and the endothelial network. SPEC control: The EC network is a primitive, capillary network. Control SPEC(6): 73.655 μm with a 3.51 μm step size (22 steps). No Rx, XRT SPEC(3): 76.027 μm with a 4 μm step size (28 steps). Rx pretreatment, full exposure SPEC(1): 88.031 μm with a 4 μm step size (23 steps).

[0239] Figures 10A-10D show fluorescence images that may illustrate the effect of pretreatment with Hiroloxamer on cells exposed to 1 Gy of radiation.

[0240] Figure 11 shows fluorescence images that may indicate that 2 Gy exposure can lead to intracellular gap formation, which can be mitigated by pretreatment with Hiroloxamer. Tension fibers are required to induce cell contraction and dramatically influence the rate and size of interendothelial gaps formed when cells retract from their boundaries (Pasain). Direct association of actin cytoskeleton with cell adhesion proteins is essential for barrier function. Tight junctions and adherent junctions connect adjacent cells and regulate paracellular permeability. Without Rx: Intercellular gap formation indicates a loss of cell-cell junctions, leading to depolymerization of F-actin in Ecs.

[0241] Figure 12 shows a fluorescence image that may demonstrate the effect of pretreatment with Hiroloxamer on cells exposed to 50 Gy of radiation.

[0242] Example 6 - Efficacy of Hiroloxamer pretreatment in vivo At least in this embodiment, the effect of pretreatment with Hiroloxamer on radiation damage to lung tissue is discussed and demonstrated. Control healthy lung tissue for comparison with the results presented herein is discussed and shown in Cho et al., Korean J Physiol Pharmacol. 2013. Aug:17(4):267-274 and Almeida et al (2013) PLoS ONE 8(1):e53628. Histological presentations of reference for the progression of radiation-induced lung injury are discussed in Sun et al. J. Radiat Res. 2014. Jul:55(4):683-689 and Almeida et al (2013) PLoS ONE 8(1):e53628. Cho et al. also discuss and provide histological presentations of macroscopic changes in the lung's features after radiation exposure, particularly thickening of the vascular lining around the bronchioles, thickening of the fine lung parenchyma, inflammatory infiltration, and deposition of fibrinous exudate, as well as vascular edema and thickening.

[0243] Figure 28 shows the irradiation protocols for these experiments whose results are shown in Figures 13 to 27. Figure 29 shows an overview of the animal groups used in these experiments.

[0244] Figure 13 shows the lung histology of PBS- or Hiroloxamer-treated rats that may show the effect of 200 mg / kg Hiroloxamer treatment 3 hours before 20Gy irradiation. Lung samples for histology were collected at about 24 hours after irradiation.

[0245] Figure 14 shows the lung histology of PBS- or Hiroloxamer-treated rats that may show the effect of 200 mg / kg Hiroloxamer treatment administered 15 minutes before exposure to 20Gy irradiation.

[0246] Figure 15 shows the lung histology of PBS- or Hiroloxamer-treated rats that may show the effect of 200 mg / kg Hiroloxamer treatment administered 15 minutes before exposure to 20Gy irradiation. The histological reference for healthy controls was obtained from http: / / histology.oucreate.com / Captions / Respiratory / 109.ling.mammal / 109.bronchiole.c1.40.Lhtm#click.

[0247] Figure 16 shows the lung histology of PBS- or Hiroloxamer-treated rats that may show the effect of 200 mg / kg Hiroloxamer treatment administered 15 minutes before exposure to 20Gy irradiation.

[0248] Figure 17 shows the lung histology of PBS- or Hiroloxamer-treated rats that may show the effect of 200 mg / kg Hiroloxamer treatment administered 15 minutes before exposure to 20Gy irradiation.

[0249] Figure 18 shows the lung histology of PBS- or Hiroloxamer-treated rats, which can demonstrate that Hiroloxamer can preserve the oxygen pathway in bronchioles in irradiated lungs. The histological reference for healthy controls was obtained from http: / / histology.oucreate.com / Captions / Respiratory / 109.ling.mammal / 109.bronchiole.c1.40.Lhtm#click.

[0250] Figure 19 shows the lung histology of PBS- or Hiroloxamer-treated rats, which can demonstrate the effect of 200 mg / kg Hiroloxamer treatment administered 15 minutes before exposure to 20 Gy irradiation.

[0251] Figure 20 shows the lung histology of PBS- or Hiroloxamer-treated rats, which can demonstrate that Hiroloxamer attenuates bronchiolar inflammation through vascular protection.

[0252] Figure 21 shows the lung histology of PBS- and Hiroloxamer-treated rats, as well as healthy controls, which can demonstrate that Hiroloxamer can preserve vascular (V) and bronchiolar (B) structures.

[0253] Figure 22 shows the lung histology of PBS- and Hiroloxamer-treated rats, which can demonstrate that Hiroloxamer can preserve vascular (V) and bronchiolar (B) structures. Figure 22 is a high-magnification image of that shown in Figure 21.

[0254] Figure 23 shows the lung histology of PBS- and Hiroloxamer-treated rats, which can demonstrate that Hiroloxamer can preserve vascular (V) and bronchiolar (B) structures. Healthy controls are also shown.

[0255] Figure 24 shows the lung histology of PBS and Hiroloxamer, which can demonstrate that Hiroloxamer can attenuate early radiation-induced injury in the lung.

[0256] Figure 25 shows lung histology from PBS and hiroloxamer-treated rats, which may demonstrate that perivascular tissue in the lungs irradiated with hiroloxamer can be preserved.

[0257] Figure 26 shows lung histology from PBS and Hiroloxamer-treated rats, which may demonstrate that Hiroloxamer can preserve the perivascular tissue of the lungs irradiated with it.

[0258] Figure 27 shows lung histology from PBS and Hiroloxamer-treated rats, which may demonstrate that Hiroloxamer can preserve the perivascular tissue of the lungs irradiated with it. Figure 27 is a higher magnification of the image shown in Figure 26.

[0259] Figure 30 shows the results one week after irradiation (20 Gy). Figure 30 shows a panel of representative micrographs of hematoxylin and eosin (H&E) stained lung sections one week after irradiation from the control, radiation (RT), and Hiroloxamer+ radiation groups (scale bar = 250 μm). The control group had normal (clear) lumens of the lung bronchi ("Br") and normal lumens of the air alveoli. The RT group clearly had inflammatory infiltration to almost obstruction throughout the entire peribronchial region, as well as collapsed blood vessels and capillaries, thickening of the alveolar septa, and cellular infiltration surrounding the bronchial structures. The Hiroloxamer+RT group had mild inflammation around the major perivascular vessels of the peribronchial region, but the alveoli and perivascular regions appeared normal.

[0260] Figure 31 shows the results 6 weeks (20 Gy) post-irradiation, which may demonstrate that Hioloxamer can preserve healthy lung tissue in a partial-volume rat lung X-ray irradiation model. Figure 31 shows representative micrographs of hematoxylin and eosin (H&E) stained right lower lung lobe sections from control, radiotherapy (RT), and Hioloxamer+radiation 6 weeks post-irradiation (scale bar = 250 μm). The control group had normal (clear) lumens of the lung bronchi ("Br") and normal lumens of the air alveoli. The RT group had marked interstitial edema, vascular and capillary ("V") congestion, increased thickness of the alveolar septa, and dense inflammatory infiltration throughout the entire lobe distally dilated in the right upper lung lobe. The Hioloxamer+RT group had some inflammatory infiltration surrounding some bronchioles located near some minor alveolar thickenings, with intact blood vessels and minimal surrounding inflammatory infiltration.

[0261] Figure 32 shows a representative micrograph of a picrosilius red-stained right lower lung lobe section 6 weeks after irradiation with 20 Gy of X-rays (scale bar = 250 microns).

[0262] Figure 33 shows representative micrographs of hematoxylin and eosin (H&E) stained right upper lung lobe sections from control, radiation (RT), and Hiroloxamer+ radiation 6 weeks after irradiation (scale bar = 250 microns). Radiation-induced lung injury spread from the radiation-exposed right lower lung lobe to the distal right upper lung lobe only in the 20 Gy RT group.

[0263] Pharmacokinetics primarily reflect renal clearance, which was calculated as follows: Clearance = 7744 mL / hour. max = 0.42 mg / hour. T max =30.49 hours, T1 / 2 = 4.77 hours, mean residence time (MRT) = 34.06 hours. Treatment period: 100-150 mg / kg loading dose over 1 hour, followed by maintenance of 20-40 mg / kg / hour. Duration of action: t 1 / 2 = 4.77 hours (The plasma half-life after infusion is approximately 4.5 hours). C max = Maximum (peak) plasma drug concentration T max=Time to reach the maximum (peak) plasma concentration after drug administration. No differences were observed between male and female patients in any subgroup or for any pharmacokinetic parameter.

[0264] Example 7 - Hiroloxamer can prevent radiation-induced pneumonia and radiotoxicity. Figure 34 shows representative images and micrographs that may demonstrate that Hiroloxamer can prevent the development of acute pneumonia in a rat model of radiation-induced pneumonia. Exposure of SD rats to a single dose of 20 Gy of X-ray irradiation is the prescription dose for radiation-induced pneumonia (Ghita). The rats were anesthetized and restrained in a custom-made jig designed to expose only 6 mm of the right central lung lobe to irradiation, while the rest of the animals remained shielded by a lead shield. To examine the development and progression of pneumonia, proximal and distal tissues of the irradiated site were stained with hematoxylin and eosin (H&E), and the integrity of major lung structures was assessed 6 weeks post-irradiation. Representative micrographs of right lower lung lobe sections from healthy normal, vehicle (saline) + RT, and Hiroloxamer + RT 6 weeks post-irradiation. Vehicle (saline) + RT control animals developed acute pneumonia within 6 weeks post-irradiation, as evidenced by capillary leakage, marked interstitial edema, vascular and capillary congestion, increased alveolar thickness, and dense inflammatory infiltration distally propagating to adjacent lobes and the contralateral left lung. Animals treated with Hiroloxamer before irradiation (Hiroloxamer + RT) had clear peribronchial and bronchial lumens and appeared normal alveolar septa, similar to healthy control animals (healthy normal); n=3 for each group.

[0265] Figures 35A–35D show representative micrographs (Figures 35A–35C) and graphs (Figure 35D) illustrating MPO analysis of leukocyte infiltration. Controlled neutrophil degranulation and myeloperoxidase (MPO) release at the injury site are necessary for effective wound healing. However, excessive degranulation can exaggerate the inflammatory response and lead to tissue damage even in the absence of infection. The level of MPO activity indicates the state of inflammation and oxidative stress in the tissue. To model radiation-induced pneumonia, SD rats were irradiated with a single 20 Gy dose of X-rays through a 6 mm well of lead shield in the right central lung lobe, as previously described. Sections from three right and contralateral left lung lobes were paraffin-embedded, sectioned to 5 μm, and stained with the Hanker-Yates peroxidase leukocyte kit (Sigma Aldrich) for analysis. Stained sections were imaged using a LionHeartFX automated microscope, and MPO+ cell counting was automated via thresholding in Gen5 software. Three animals were used in each group, and five high-power areas were analyzed for each animal. (Figure 35A) Representative MPO-stained right lower lung lobe sections of healthy controls, (Figure 35B) vehicle (saline) + RT, and (Figure 35C) 200 mg / kg Hiroloxamer + RT. Tissue damage was observed in the control saline + RT animals, and they had significantly elevated MPO levels in all lung lobes compared to Hiroloxamer + RT and healthy normal animals (Figure 35D). Statistical comparisons (ANOVA, followed by Tukey) of healthy normal, vehicle (saline) + RT, and hiroloxamer + RT showed no difference between healthy normal and hiroloxamer + RT, while significant statistical differences were found between vehicle (saline) + RT and healthy normal (p<0.001), and between vehicle (saline) + RT and hiroloxamer + RT (p<0.001).

[0266] Figure 37 shows representative microscopic images that may demonstrate that Hiroloxamer protects healthy tissue from radiotoxicity, regardless of radiation dose. Animals were irradiated with either 10 Gy × 1 fx, 20 Gy × 1 fx, or 10 Gy × 4 fx X-rays at 2-3 days intervals for 2 weeks in an 8 mm area of ​​the right lung. All animals were sacrificed 6 weeks after irradiation and stained with H&E (5 μm sections). Animals treated with the vehicle (upper panel of the figure) had varying degrees of apparent radiation-induced lung injury based on the dose. The 20 Gy × 1 fx dose resulted in the most significant injury, which was evidenced by edema, inflammatory infiltration, and congestion of the lung parenchyma, which were not localized to the irradiation but extended to all lobes on both sides. These histological findings indicate the development of radiation-induced acute pneumonia. 10 Gy × 1 fx and 10 Gy × 4 fx caused less damage than 20 Gy × 1 fx, which is consistent with the classical theory that healthy tissue toxicity depends on the dose per dose rather than the total dose delivered. Animals pre-treated with 200 mg / kg of Hiroloxamer showed only localized damage in the targeted irradiation area, and healthy tissue was protected from toxicity in all radiotherapy regimens evaluated.

[0267] Figures 38A–38D show representative microscopic images that may demonstrate dose-dependent protection of healthy tissue from radiotoxicity by hiroloxamer. Animals were exposed to 10 Gy x 4 doses of X-ray irradiation delivered every 2–3 days over two weeks in an 8 mm area of ​​the right lung. Animals treated with 200 mg / kg of hiroloxamer before irradiation (Figure 38D) had a normal lung appearance comparable to that of healthy controls (Figure 38A). Animals treated with 50 mg / kg of hiroloxamer (Figure 38C) had evident cellular infiltration and subsequent alveolar thickening, which was not as severe as in vehicle (PBS) treated animals (Figure 38B), but did not provide adequate protection compared to the 200 mg / kg dose.

[0268] Various modifications and variations of the described methods, pharmaceutical compositions, and kits of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been described in connection with specific embodiments, it will be understood that further modifications are possible and the claimed invention should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in the art are intended to be within the scope of the present invention. This application is intended to generally embrace any variations, uses, or adaptations of the present invention in accordance with the principles of the present invention, including such departures from the present disclosure as come within known practice in the art to which the invention pertains and can be applied to the essential features described hereinabove.

[0269] Further attributes, features, and embodiments of the present invention can be understood by reference to the following numbered aspects of the disclosed invention. References to the disclosure in any of the preceding aspects are applicable to any preceding numbered aspect, and any combination of any number of preceding aspects, as may be recognized by appropriate preceding disclosure in any combination of preceding aspects that may be made. The following numbered aspects are provided.

[0270] 1. A method for preventing damage to non-injured cells in a subject during and / or after injury exposure, comprising: administering a first amount of poloxamer 188 (P188) or a pharmaceutical formulation thereof to the subject before exposing one or more regions of the subject to injury; exposing one or more regions of the subject to injury after the blood concentration of P188 reaches 1 - 5 mg / mL; and.

[0271] 2. The method of aspect 1, wherein the first amount of P188 is administered intravenously.

[0272] 3. n thAny one of embodiments 1 to 2 further comprises administering to a subject an amount of P188 or a pharmaceutical preparation thereof after administering a first amount of P188 and before exposing one or more areas of the subject to injury, wherein n is 2 or more.

[0273] 4. The first quantity on page 188, n th One of the methods in any of embodiments 1 to 3, wherein the amount, or the total amount, is an amount effective in raising the blood concentration of P188 to 1 to 5 mg / mL within 1 to 48 hours.

[0274] 5. The first quantity or n on page 188 th A method in which the amount is administered as a continuous infusion over a period of time, in any one of embodiments 1 to 4.

[0275] 6. The first quantity or n on page 188 th One of the methods described in any of embodiments 1 to 5, wherein one or more of the amounts are administered as a bolus dose.

[0276] 7. Any one of embodiments 1-2 and 4-6, wherein only a first dose is administered, and the first dose is effective in raising the blood concentration of P188 to approximately 1-5 mg / mL within 1-48 hours.

[0277] 8. The subject is the first or n on page 188. th One of the methods in embodiments 1 to 7, wherein after receiving the amount of P188, the amount of P188 is not received for at least 14 hours.

[0278] 9. Any one of embodiments 1 to 8, wherein P188 administration is discontinued immediately after exposure of one or more areas of the target to injury.

[0279] 10. Exposure of one or more areas of the subject to injury constitutes the first or n th One of the methods described in embodiments 1 to 9, performed approximately 1 to 48 hours after administration of P188.

[0280] 11. Any one of the methods described in 1 to 10, wherein the injury is effective in killing one or more cancerous cells in one or more regions of a target.

[0281] 12. Any one of embodiments 1 to 11, wherein the non-injured cells are non-cancerous cells.

[0282] 13. Any one of embodiments 1 to 12, wherein the non-injured cells are endothelial cells.

[0283] 14. Any one of embodiments 1 to 13, wherein the injury is a mechanical injury, chemical injury, biological injury, energetic injury, physiological injury, or a combination thereof.

[0284] 15. Any one of the methods described in aspects 1 to 14, wherein the injury is caused by ionizing radiation.

[0285] 16. A method for preventing damage to non-injured cells in a subject during and / or after injury exposure, comprising: Administering a certain amount of poloxamer 188 (P188) or a pharmaceutical preparation thereof to a subject before exposure of one or more areas of the subject to injury, wherein the amount is in the range of approximately 10 to 150 mg / kg; Exposure to one or more areas of the target area for injury within 0 to 48 hours after administration of a certain amount of P188 or a pharmaceutical preparation thereof, Methods that include...

[0286] 17. The method according to embodiment 16, wherein a certain amount of P188 or a pharmaceutical preparation thereof is administered intravenously.

[0287] 18. Any one of the methods described in 16 to 17, wherein the administration of P188 or its pharmaceutical preparation is discontinued immediately before or immediately after exposure of one or more areas of the target to injury.

[0288] 19. Any one of the methods described in 16 to 18, wherein the subject receives no further dose of P188 or its pharmaceutical preparation for at least 14 hours after being administered a certain dose of P188 or its pharmaceutical preparation, or after discontinuing the administration of P188 or its pharmaceutical preparation.

[0289] 20. Any one of the methods described in embodiments 16 to 18, wherein the injury is a mechanical injury, a chemical injury, a biological injury, an energetic injury, a physiological injury, or a combination thereof.

[0290] 21. Any one of the methods described in aspects 16 to 20, wherein the injury is caused by ionizing radiation.

[0291] twenty two. A certain amount of P188 or a pharmaceutical formulation thereof, which is effective in preventing damage to non-injured cells in a subject when one or more areas of the subject are exposed to injury; Instructions, provided in a tangible medium, to administer a certain amount of P188 or its pharmaceutical formulation to a subject 0 to 48 hours before, 0 to 48 hours after, or both of, exposure to an injury in one or more areas of the subject, and A kit that includes this.

[0292] 23. A kit according to embodiment 22, wherein the injury is mechanical, chemical, biological, energetic, physiological, or a combination thereof.

[0293] 24. One of the kits according to any one of embodiments 22-23, wherein the injury is caused by ionizing radiation.

[0294] 25. Any one of the kits in any of embodiments 22 to 24, wherein the instructions further instruct the subject to discontinue administration of a certain amount of P188 or its pharmaceutical formulation immediately before or after exposure to injury to one or more areas of the subject.

[0295] 26. Any one of the kits described in embodiments 22 to 25, wherein a certain amount of P188 or a pharmaceutical formulation thereof is effective in achieving a blood concentration of approximately 1 to approximately 5 mg / mL in a subject within 0 to 48 hours.

[0296] 27. A method for protecting non-injured cells after exposure to an injury target, the following: a) Administering a certain amount of P188 or a preparation thereof to the subject before exposure to injury; b) Administering a certain amount of P188 or a preparation thereof to the subject immediately after exposure to injury; c) Administering a certain amount of P188 or a preparation thereof to the subject during injury; or d) combinations of those Methods that include...

[0297] 28. The method of embodiment 27, wherein a certain amount is effective in increasing the blood concentration of P188 in a subject to 1 mg / mL to 5 mg / mL.

[0298] 29. Any one of the methods described in embodiments 27 to 28, wherein a certain amount is effective in coating one or more non-injured cells.

[0299] 30. Any one of embodiments 27 to 29, wherein a certain amount is effective in reducing or preventing oxidative damage to one or more non-injured cells.

[0300] 31. Any one of embodiments 27 to 30, wherein a certain amount is effective in reducing or preventing inflammatory damage to one or more non-injured cells.

[0301] 32. Any one of embodiments 27 to 31, wherein the administration is carried out 0 to 48 hours before exposure to injury.

[0302] 33. Any one of the methods described in embodiments 27 to 32, wherein the administration is performed 0 to 48 hours or 0 to 24 hours after exposure to injury.

[0303] 34. Any one of the methods in any of embodiments 27 to 32, wherein the injury is a mechanical injury, a chemical injury, a biological injury, an energetic injury, a physiological injury, or a combination thereof.

[0304] 35. Any one of the methods described in aspects 27 to 34, wherein the injury is caused by ionizing radiation.

[0305] 36. A method for preventing injury-induced pneumonia in a subject, wherein the method is: a) Administering a certain amount of P188 or a preparation thereof to the subject before exposure to injury; b) Administering a certain amount of P188 or a preparation thereof to the subject immediately after exposure to injury; c) Administering a certain amount of P188 or a preparation thereof to the subject during injury; or d) combinations of those Methods that include...

[0306] 37. The method of embodiment 36, wherein a certain amount is effective in increasing the blood concentration of P188 in a subject to 1 mg / mL to 5 mg / mL.

[0307] 38. Any one of embodiments 36 to 37, wherein a certain amount is effective in coating one or more non-injured cells.

[0308] 39. Any one of embodiments 36 to 38, wherein a certain amount is effective in reducing or preventing oxidative damage to one or more non-injured cells.

[0309] 40. Any one of embodiments 36 to 39, wherein a certain amount is effective in reducing or preventing inflammatory damage to one or more non-injured cells.

[0310] 41. Any one of embodiments 36 to 40, wherein the administration is carried out 0 to 48 hours before exposure to injury.

[0311] 42. Any one of the methods described in embodiments 36 to 41, wherein the administration is performed 0 to 24 hours or 0 to 48 hours after exposure to injury.

[0312] 43. Any one of the methods in any of embodiments 36 to 42, wherein the injury is a mechanical injury, a chemical injury, a biological injury, an energetic injury, a physiological injury, or a combination thereof.

[0313] 44. Any one of the methods described in aspects 36 to 43, wherein the injury is caused by ionizing radiation.

[0314] 45. A method for treating a disease or disorder in a person requiring treatment for a disease or disorder, wherein the method is: a) Administering a certain amount of P188 or a preparation thereof to the subject before exposure to injury; b) Administering a certain amount of P188 or a preparation thereof to the subject immediately after exposure to injury; c) Administering a certain amount of P188 or a preparation thereof to the subject during injury; or d) combinations of those Methods that include...

[0315] 46. ​​The method of aspect 45, wherein a certain amount is effective in increasing the blood concentration of P188 in a subject to 1 mg / mL to 5 mg / mL.

[0316] 47. Any one of embodiments 45 to 46, wherein a certain amount is effective in coating one or more non-injured cells.

[0317] 48. Any one of embodiments 45 to 47, wherein a certain amount is effective in reducing or preventing oxidative damage to one or more non-injured cells.

[0318] 49. Any one of embodiments 45 to 48, wherein a certain amount is effective in reducing or preventing inflammatory damage to one or more non-injured cells.

[0319] 50. Any one of embodiments 45 to 49, wherein the administration is carried out 0 to 48 hours after exposure to injury.

[0320] 51. Any one of the methods described in aspects 45 to 50, wherein the administration is carried out 0 to 24 hours or 0 to 48 hours after exposure to injury.

[0321] 52. Any one of the methods described in aspects 45 to 51, wherein the injury is a mechanical injury, a chemical injury, a biological injury, an energetic injury, a physiological injury, or a combination thereof.

[0322] 53. The method of aspect 52, wherein the injury is an ischemic event, possibly a stroke or myocardial infarction.

Claims

1. A polixamer 188 (P188) or a pharmaceutical formulation thereof, for preventing damage to undamaged cells from injury-induced free radicals or inflammatory cascades produced from directly damaged cells in a subject during and / or after injury exposure, A first dose of poloxamer 188 (P188) or a pharmaceutical formulation thereof is administered to one or more areas of the subject before they are exposed to injury; or, after the blood concentration of P188 reaches 1 to 5 mg / mL, one or more areas of the subject are exposed to injury, A first dose of poloxamer 188 (P188) or a pharmaceutical formulation thereof is administered to one or more areas of the subject before exposure of those areas to injury; the dose is in the range of approximately 10 to 150 mg / kg; and within 0 to 48 hours after administration of the dose of P188 or a pharmaceutical formulation thereof, one or more areas of the subject are exposed to injury. Agent.

2. (a) A first dose of P188 or its pharmaceutical preparation is administered intravenously. (b) A first dose or nth dose of P188 or its pharmaceutical formulation is administered as a continuous infusion over a period of time. (c) One or more of the first amount or the amount of nth, P188 or its pharmaceutical preparation, is administered as a bolus dose. (a) and (b), or (a) and (c) That is, The agent according to claim 1.

3. The agent according to claim 1, wherein an amount of nth of P188 or a pharmaceutical formulation thereof is further administered to the subject after administering the first amount of P188 or a pharmaceutical formulation thereof, and before one or more areas of the subject are exposed to the injury, wherein n is 2 or more.

4. The agent according to claim 1, wherein the first amount of P188, the amount of nth, or the total amount is an amount effective in raising the blood concentration of P188 to 1 to 5 mg / mL within 1 to 48 hours.

5. The agent according to claim 1, wherein the subject receives no further dose of P188 or its pharmaceutical preparation for at least 14 hours after receiving the first or nth dose of P188 or its pharmaceutical preparation.

6. The agent according to claim 1, wherein the administration of P188 or the pharmaceutical preparation thereof is discontinued immediately after one or more areas of the subject are exposed to the injury.

7. The agent according to claim 6, wherein the subject does not receive any amount of P188 or its pharmaceutical preparation for at least 14 hours after discontinuing administration of P188 or its pharmaceutical preparation.

8. The agent according to claim 1, wherein exposure of one or more areas of the subject to the injury is performed approximately 1 to approximately 48 hours after administration of the first or nth P188 or the pharmaceutical formulation thereof.

9. The agent according to claim 1, wherein the injury is effective in killing one or more cancerous cells within one or more regions of the target.

10. The agent according to claim 1, wherein the non-injured cells are non-cancerous cells, or the non-injured cells are endothelial cells, or both.

11. The agent according to claim 1, wherein the injury is a mechanical injury, a chemical injury, a biological injury, an energetic injury, a physiological injury, or a combination thereof.

12. The agent according to claim 1, wherein the injury is caused by ionizing radiation.

13. (a) an agent for protecting non-injured cells from injury-induced free radicals or inflammatory cascades produced from directly injured cells after exposure of the subject to injury; (b) an agent for preventing injury-induced pneumonia in the subject; (c) an agent for treating a disease or injury in the subject; or (d) an agent for any combination of (a) to (c), Poloxamer 188 (P188) or a pharmaceutical preparation thereof, a) A certain amount of poloxamer 188 (P188) or a pharmaceutical formulation thereof is administered to the subject before exposure to injury; b) A certain amount of P188 or a pharmaceutical formulation thereof is administered to the subject immediately after exposure to injury; c) A certain amount of P188 or a pharmaceutical formulation thereof is administered to the subject during the injury; or d) Any combination of a) to c) It is a drug.

14. The agent according to claim 13, wherein a certain amount of poloxamer 188 (P188) or a pharmaceutical preparation thereof is effective in increasing the blood concentration of P188 in the subject to 1 mg / mL to 5 mg / mL.

15. The agent according to claim 13, wherein the amount is (a) effective in coating one or more non-harmful cells, (b) effective in reducing or preventing oxidative damage to one or more non-harmful cells, (c) effective in reducing or preventing inflammatory damage to one or more non-harmful cells, or (d) any combination of (a) to (c).

16. The agent according to claim 13, wherein the administration is carried out 0 to 48 hours before exposure to the injury.

17. The agent according to claim 13, wherein administration is performed 0 to 48 hours or 0 to 24 hours after exposure to the injury.

18. The agent according to claim 13, wherein the injury is a mechanical injury, a chemical injury, a biological injury, an energetic injury, a physiological injury, or a combination thereof.

19. The agent according to claim 13, wherein the injury is caused by ionizing radiation.

20. The agent according to claim 13, wherein the injury is an ischemic event, and in some cases a stroke or myocardial infarction.

21. A certain amount of poloxamer 188 (P188) or a pharmaceutical formulation thereof, which is effective in preventing damage to undamaged cells from injury-induced free radicals or inflammatory cascades produced by directly injured cells in a subject when one or more areas of the subject are exposed to injury; Instructions, fixed in a tangible medium, that instruct the administration of a certain amount of P188 or its pharmaceutical formulation to the subject 0 to 48 hours before exposure to an injury in one or more areas of the subject, 0 to 48 hours after exposure, or both, and A kit that includes this.

22. The kit according to claim 21, wherein the injury is a mechanical injury, a chemical injury, a biological injury, an energetic injury, a physiological injury, or a combination thereof.

23. The kit according to claim 21, wherein the injury is caused by ionizing radiation, or ischemic events, optionally stroke or myocardial infarction.

24. The kit according to claim 21, wherein the instructions further indicate that administration of a certain amount of P188 or the pharmaceutical formulation thereof to one or more areas of the subject should be discontinued immediately before or after exposure to the injury.

25. The kit according to claim 21, wherein a certain amount of P188 or a pharmaceutical formulation thereof is effective in achieving a blood concentration of about 1 to about 5 mg / mL in the subject within 0 to 48 hours.

26. Use of poloxamer 188 (P188) for (a) to protect non-injured cells from injury-induced free radicals or inflammatory cascades produced from directly injured cells after exposure of the subject to injury; (b) to prevent injury-induced pneumonia in the subject; (c) to treat disease or injury in the subject; (d) to prevent damage to non-injured cells from injury-induced free radicals or inflammatory cascades produced from directly injured cells in the subject during and / or after exposure to injury; or (e) for the manufacture of agents for any combination of (a) to (d).

27. A pharmaceutical composition for use for any combination of (a) to (d), comprising poloxamer 188 (P188), wherein the composition comprises poloxamer 188 (P188), to (a) protect non-injured cells from injury-induced free radicals or inflammatory cascades produced from directly injured cells after exposure to injury of the subject, (b) prevent injury-induced pneumonia in the subject, (c) treat a disease or injury in the subject, (d) prevent damage to non-injured cells from injury-induced free radicals or inflammatory cascades produced from directly injured cells in the subject during and / or after exposure to injury, or (e) any combination of (a) to (d).

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