Gel proteins and opioid tolerance

Abstract

This invention relates to compositions and methods for treating opioid tolerance in subjects, and to methods for selecting pain relief treatment regimens for subjects suffering from and / or at risk of developing opioid tolerance. Embodiments of the treatment methods of this invention include administering a gelling agent to produce a therapeutic effect against opioid tolerance in the subject.

Description

[0001] Related applications

[0002] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 547,801, filed November 8, 2023, pursuant to 35 USC § 119(e), the disclosure of which is incorporated herein by reference in its entirety.

[0003] Government interests

[0004] This invention was made with government support under NS122855 granted by the National Institutes of Health and N00014-22-1-2818 granted by the US Office of Naval Research. The government holds certain rights in this invention. Technical Field

[0005] In some aspects, the present invention relates to compositions and methods for reducing opioid tolerance. Background Technology

[0006] Opioid tolerance is widely recognized as a serious problem in healthcare. There are currently no effective methods to reduce and / or eliminate opioid tolerance, and there is a widespread need for such methods as adjunctive therapy for moderate to severe pain. Summary of the Invention

[0007] According to one aspect of the invention, a method for treating opioid tolerance in a subject is provided, the method comprising: administering a composition comprising a gelsolin agent to the subject requiring such treatment in an amount effective in treating opioid tolerance. In some embodiments, the opioid tolerance is morphine tolerance. In some embodiments, the gelsolin agent comprises a gelsolin molecule, a functional fragment thereof, or a functional derivative of a gelsolin molecule. In some embodiments, the gelsolin molecule is plasma gelsolin (pGSN). In some embodiments, the gelsolin molecule is a recombinant gelsolin molecule. In some embodiments, the composition comprising the gelsolin agent is administered orally, sublingually, intraorally, intranasally, intravenously, intramuscularly, intrathecally, intraperitoneally, subcutaneously, intradermally, topically, rectally, vaginally, intrasynovially, or intraocularly. Administration may be via oral, sublingual, intravenous, subcutaneous, topically, intrathecal, or by inhalation. In some embodiments, administration of a gelling agent has a greater therapeutic effect on opioid tolerance in subjects compared to a control treatment effect. In some embodiments, the control treatment effect is equal to the effect on opioid tolerance in subjects without the administration of a gelling agent. In some embodiments, administration of a gelling agent reduces opioid tolerance in subjects by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to the severity of opioid tolerance in a control without the administration of a gelling agent. In some embodiments, the subjects are mammals. In some embodiments, the mammal is a human. In some embodiments, the gelling agent is administered to the subjects 1, 2, 3, 4, 5, 6, 7, 8, or more times. In some embodiments, application of a gelling agent increases the analgesic efficacy of the opioid in a subject by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to the analgesic efficacy of a control without gelling agent. In some embodiments, the method further includes administering the opioid to a subject receiving a composition comprising a gelling agent. In some embodiments, the applied gelling agent reduces the level of opioid tolerance in the subject. In some embodiments, the applied gelling agent prevents opioid tolerance in the subject. In some embodiments, the opioid includes oxycodone, fentanyl, morphine, opium, hydromorphone, hydroxymorphone, methadone, or hydrocodone.

[0008] According to one aspect of the invention, a method is provided for selecting a pain relief treatment for a subject suffering from opioid tolerance, the method comprising: (a) selecting a gelling agent treatment regimen for the subject, comprising administering a composition comprising a gelling agent in an amount effective in treating opioid tolerance in the subject; and (b) selecting a pain relief treatment regimen for the subject, comprising administering one or more opioids and / or different pain relievers to the subject. In some embodiments, the method further comprises identifying the opioids and optionally selecting one or more opioids and / or different pain relievers based at least in part on the identification of the opioids. In some embodiments, the method further comprises administering the selected pain relief treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8 or more times. In some embodiments, the method further comprises administering the selected gelling agent treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8 or more times. In some embodiments, the administration of the gellingin treatment regimen and the pain relief treatment regimen includes administering the gellingin agent before, simultaneously with, and after administration of one or more opioids and / or different pain relievers to the subject. In some embodiments, the gellingin agent comprises a gellingin molecule, wherein the gellingin molecule is optionally a recombinant gellingin molecule, and optionally a plasma gellingin (pGSN). In some embodiments, the administration of the selected gellingin treatment regimen and the selected pain relief treatment regimen has a greater therapeutic effect against opioid tolerance in the subject compared to the control treatment effect against opioid tolerance; optionally, the control treatment effect is the effect against opioid tolerance in subjects who received the selected pain relief treatment regimen but did not receive the selected gellingin treatment regimen. In some embodiments, the application of a gelling agent reduces opioid tolerance in subjects by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to a control opioid tolerance without the application of a gelling agent. In some embodiments, opioid tolerance is measured based on one or more of the following: the presence of one or more opioid tolerance symptoms in the subject and the severity of opioid tolerance in the subject. In some embodiments, the subject is a mammal; optionally, the subject is a human.In some embodiments, the application of a gelling agent increases the pain-relieving efficacy of the selected pain-relieving treatment regimen by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, 200%, or more compared to a control group without the application of a gelling agent. In some embodiments, the opioid includes oxycodone, fentanyl, morphine, hydromorphone, methadone, or hydrocodone.

[0009] According to another aspect of the invention, a method is provided for selecting a pain relief treatment for a subject at risk of developing opioid tolerance, the method comprising: (a) selecting a gelling agent treatment regimen for the subject, comprising administering a composition comprising a gelling agent in an amount effective in reducing the risk of opioid tolerance in the subject; and (b) selecting a pain relief treatment regimen for the subject, comprising administering one or more opioids and / or different pain relievers to the subject. In some embodiments, the method further comprises administering the selected pain relief treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8 or more times. In some embodiments, the method further comprises administering the selected gelling agent treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8 or more times. In some embodiments, administering the gelling agent treatment regimen and the pain relief treatment regimen comprises administering the gelling agent to the subject one or more times before, simultaneously with, and after administering one or more opioids and / or different pain relievers. In some embodiments, the coagulant agent comprises a coagulant molecule, wherein the coagulant molecule is optionally a recombinant coagulant molecule. In some embodiments, the coagulant molecule is plasma coagulant protein (pGSN). In some embodiments, administration of a coagulant treatment regimen and a pain relief treatment regimen reduces the risk of opioid tolerance in subjects compared to a control treatment effect against the risk of opioid tolerance, optionally wherein the control treatment effect is equal to the effect against opioid tolerance in subjects who received the pain relief treatment regimen but did not receive the coagulant treatment regimen. In some embodiments, administration of a coagulant agent reduces the risk of opioid tolerance in subjects by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to opioid tolerance in a control without coagulant agent administration. In some embodiments, the subject is a mammal, optionally, a human. In some embodiments, the application of a gelling agent increases the pain-relieving efficacy of the selected pain-relieving treatment regimen in the subject by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, 200%, or more compared to the pain-relieving efficacy of the selected pain-relieving treatment regimen in a control group without the application of a gelling agent. In some embodiments, the opioid includes oxycodone, fentanyl, morphine, hydromorphone, methadone, or hydrocodone.

[0010] According to another aspect of the invention, a pharmaceutical composition comprising a gelling agent and an opioid is provided. In some embodiments, the gelling agent comprises a gelling protein molecule, wherein the gelling protein molecule is optionally a recombinant gelling protein molecule, and optionally a plasma gelling protein (pGSN). In some embodiments, the opioid is morphine, heroin, hydromorphone, hydroxymorphone, fentanyl, methadone, oxycodone, hydrocodone, codeine, buprenorphine, dextromethorphan, dextropropoxyphene, loperamide, meperidine, opioid, carfentanyl / carfentanil, morphone, talpentadol, tramadol, or pethidine. Attached Figure Description

[0011] These and other features, aspects, and advantages of embodiments of this disclosure can be better understood when the following detailed description is read with reference to the accompanying drawings, in which the same reference numerals denote the same parts throughout the drawings:

[0012] Figure 1 This is a schematic diagram illustrating the interpretation of data from morphine tolerance studies. The diagram illustrates the circulatory nature of inflammation triggered by multiple stimuli, in which astrocytes and microglia produce microparticles (MPs) that travel through the glymphatic system. These MPs travel from the perivascular pathway of the central nervous system to the deep cervical lymph nodes and subsequently into the systemic circulation. CNS-derived MPs from astrocytes that express platelet response protein-1 induce neutrophil activation and the production of additional MPs, some expressing filamentous (F-) actin. F-actin MPs lead to auto-activation of neutrophils, exacerbating the inflammatory process, and neutrophils adhering to cerebral microvasculature trigger further astrocyte and microglia activation. F-actin MPs can be disrupted by plasma coagulants (pGSN), and when this occurs, the concentration of pGSN in the bloodstream decreases.

[0013] Figure 2A To B, diagrams and charts are provided. Figure 2A This is a schematic diagram illustrating the interpretation of some research findings, demonstrating that brain-derived melanoma can initiate a cyclical neuroinflammatory process. This interpretation differs from... Figure 1This study provides further details on why F-actin MPs exhibit pro-inflammatory properties. In these other studies, F-actin positive granules have now been shown to contain high concentrations of nitric oxide synthase 2 (NOS2, the inflammatory form of nitric oxide synthase) and interleukin-1β. Plasma coagulants disrupt these MPs, and in the process, plasma coagulant levels decrease. Figure 2B A graph shows the experimental results of reduced plasma coagulants in morphine-tolerant mice and mice injected with F-actin-positive MP obtained from morphine-tolerant mice.

[0014] Figures 3A to 3B provide schematic diagrams and lists illustrating the following: Retraction test protocol (Figure 3A), which involves inducing morphine tolerance in mice by injecting morphine three times daily and testing on day 4. Figure 3B This is a graph showing the time required for mice to remove their tails from warm water, measured after a cumulative dose of morphine administered at baseline (without morphine injection) and subsequently via intraperitoneal (IP) injections of 0.5, 1, 2, and 3 mg / kg. The results indicate that controls showed a slower response in removing their tails due to the arbitrary doses (0.5 to 3 mg / kg) of morphine, while morphine-tolerant mice responded to the warm water by removing their tails despite morphine injections (0.5, 1, or 2 mg / kg).

[0015] Figure 4 The following diagrams and graphs are provided to illustrate the direction of CSF flow in the brain: the top graph shows the direction of CSF flow in the brain; the bottom left graph shows the lymphatic system in the mouse brain; and the bottom right graph is a graph of particle concentrations in controls and morphine-tolerant subjects. This graph illustrates the glial lymphatic system and the conventional lymphatic system. Fluid from both flows to the deep cervical lymph nodes. Studies measuring particle (MP) levels in the deep cervical lymph nodes were conducted, and results showed significantly elevated MP levels in morphine-tolerant mice (shown in the graph at the bottom right).

[0016] Figure 5A graph showing the number of microparticles present in the deep cervical lymph nodes of mice under morphine tolerance and under tolerance regimens (see Figure 3) with the addition of multiple drugs to antagonize different inflammatory pathways is provided. These include: PEGylation polymer B (0.7 μl of 0.3% solution [w / v] IV / g) [PEGylation polymer B is an agent that destroys most MPs when injected into the bloodstream], anakinra (100 mg / kg IP) [anakinra is a drug that blocks the receptors of interleukin-1β], methylnaltrexone (1 mg / kg IP) [methylnaltrexone is an opioid receptor blocker that does not cross the blood-brain barrier], or neutropenic animals [mice treated with antibodies to selectively remove polymorphonuclear leukocytes (also known as neutrophils) from the bloodstream]. Data are presented as mean ± SE, and numbers represent the number of mice in each group. The letters on the right list the protein markers studied on a single MP and include Ly6 (mature neutrophils), CD45 (all leukocytes), P2Y12 and CD45 (microglia), CD41 (platelets), CD31+ / CD41-dim (endothelium), glial fibrillary acidic protein GFAP (astroglia), myelin basic protein MBP (oligodendrocytes), or neuronal pentamerin receptor NPR (neurons).

[0017] Figure 6 This provides evidence of tolerance antagonism and demonstrates that treatment of mice with an agent that inhibits particle elevation (as shown in Figure 3) can prevent anti-nociceptive tolerance (e.g.) Figure 1 The figure shown is shown in the image. Figure 6 The time required for mice to remove their tails from warm water, measured at baseline and subsequently after cumulative morphine doses of 0.5, 1, 2, and 3 mg / kg IP, is shown. Studies were conducted after daily administration of sterile saline (control) or morphine for two consecutive days at 20 mg / kg followed by 50 mg / kg on day 3. It was indicated that mice also received anaspirin (100 mg / kg IP), methylnaltrexone (1 mg / kg IP), or PEGylation B (0.7 μl of 0.3% solution [w / v] IV / g). Data are mean ± SE of repeated measures, p < 0.05, two-way ANOVA.

[0018] Figure 7Western blot images of the precursor and mature forms of IL-1β are provided. Plasma samples containing 100,000 MP were centrifuged at 21,000 × g for 1 hour. The precipitate was retained for Western blotting, and the supernatant was centrifuged at 100,000 g for 1 hour so that the precipitate could also be Western blotted to assess the relative amounts of precursor and mature IL-1β. The data below the figure show the concentrations of IL-1β, in pg / 1,000,000 MP, from whole plasma and the supernatant and precipitate fractions after 21,000 g centrifugation. Data are mean ± SE, n = 6. The results shown in the table indicate that IL-1β is elevated in tolerant mice and that almost all of IL-1β is present in the microparticles. The results show that no IL-1β was present in the 100,000 g plasma supernatant (therefore packaged in vesicles).

[0019] Figure 8 A table showing results from the following study: blood samples were collected from patients newly admitted to methadone treatment for opioid use disorder and age-matched, medication-naïve controls. Data are presented as mean ± SD, n=40 per group. The results, using a t-test, showed that blood-derived MP levels were elevated in patients receiving methadone treatment compared to control levels.

[0020] Figures 9A to 9C provide photographs and graphs illustrating measurements of glial lymphatic flow assessment performed by gadolinium (Gd)-contrast MRI. Figure 9A highlights the signal near the Galen vein and deep cervical lymph nodes. Flow was quantified at these sites using a 1 mm target region, and the signal was normalized by including a 60 mM Gd vial under the mouse skull within the scanning field. Figure 9B Images of control and tolerant mice are shown in the image. Figure 9C The figure shows a graph of quantified glial lymphatic flow measured in 5 mice. The vertical axis represents the normalized Gd signal, and the horizontal axis represents the time following Gd injection in the mice. The MRI machine was programmed to collect images for 5 minutes and then report the values. Therefore, the horizontal axis is shown at consecutive 5-minute intervals. The data indicate that morphine-tolerant mice exhibited increased glial lymphatic flow compared to control mice that received only sterile saline.

[0021] Figure 10Schematic diagrams, graphs, and Western blot images illustrating experiments conducted by injecting F-actin-positive MP from morphine-tolerant mice into naïve mice are provided. Overall, the results indicate that morphine injection increases glial lymphatic flow, and this contributes to the elevation of MP present in the deep cervical lymph nodes and blood. Results from the PEG-modifier B injection study suggest that blood-derived MP is required for morphine tolerance. Further research indicates that, in particular, F-actin-positive MP triggers persistent inflammation. Figure 10 The images show that when mice were injected with F-actin MP from tolerant mice, they exhibited somewhat similar increases in glial lymphatic flow (left panel) and neuroinflammation (right Western blot). The numbers under each band represent the mean ± SD (n = 3 brains / lane) band density relative to control samples from a repeat study. Western blots probed IBA-1 (which indicates microglia activation), aquaporin-4 (AQP4, a protein that drives glial lymphatic flow in astrocytes), myeloperoxidase (MPO, evidence of neutrophil adhesion to the cerebral vascular system), and CD36 protein (which, when elevated, is associated with neuroinflammation).

[0022] Brief description of the sequence

[0023] SEQ ID NO: 1 is a GenBank ® The amino acid sequence of human plasma coagulation protein from login number X04412:

[0024] Detailed Implementation

[0025] This invention is partly based on the unexpected discovery that gelling agents can be applied to subjects to treat opioid tolerance. Certain embodiments of the method of this invention include a treatment regimen comprising administering a therapeutic composition containing a gelling agent to a subject identified as having opioid tolerance or at risk of developing opioid tolerance. Certain aspects of the invention include selecting pain-relieving treatments for subjects known to have opioid tolerance or at risk of developing opioid tolerance. Certain embodiments of the method of this invention can improve the efficacy of pain management in subjects by applying gelling agents. It has now been determined that increasing plasma gelling agent (pGSN) levels in subjects receiving opioid agents can reduce opioid tolerance in the subjects.

[0026] Opioids and Opioid Tolerance

[0027] Opioids are widely used analgesics to provide relief from moderate to severe pain, but their use in chronic pain relief or other pain conditions can lead to a decrease in their potency. Opioid tolerance is the physical effect of repeated opioid use and can lead to opioid-induced hyperalgesia, also described as increased pain with a gradual increase in opioid dosage. Opioid-induced hyperalgesia is a condition of nociceptive sensitization caused by exposure to opioids. This condition is characterized by a paradoxical response in which patients receiving opioids for pain treatment may actually become more sensitive to certain painful stimuli. Hyperalgesia can manifest as opioid-induced tolerance because increased sensitivity to pain counteracts the analgesic effect of opioids. Subjects may develop opioid tolerance in response to persistent and / or repeated opioid use, and subjects with opioid tolerance may exhibit reduced pain relief in response to opioid medications. Opioid tolerance can arise from the administration of illicit and / or legal opioids to subjects.

[0028] Many opioids are used for pain relief in healthcare and are also used illicitly. The term "opioid" as used herein may refer to both natural and synthetic opioids. Some non-limiting examples of opioids that can induce tolerance are morphine, heroin, hydromorphone, hydroxymorphone, fentanyl, methadone, oxycodone, hydrocodone, codeine, buprenorphine, dextromethorphan, dextropropoxyphene, loperamide, meperidine, opioids, carfentanil / carfentanil, morphone, tapentadone, tramadol, and meperidine.

[0029] The loss of therapeutic pain relief benefits from opioids due to ongoing or prior opioid use is termed "opioid tolerance," which can lead to the need to increase the amount and dosage of opioids administered in individuals with opioid tolerance to maintain satisfactory levels of pain relief. Furthermore, managing new moderate or severe pain (e.g., pain resulting from surgery or injury) can be extremely difficult for patients with opioid tolerance caused by opioid treatment for conditions (e.g., but not limited to chronic pain) or by illicit opioid use.

[0030] The FDA defines opioid tolerance in human subjects as being caused by administration of "at least 60 mg oral morphine / day, 25 mcg transdermal fentanyl / hour, 30 mg oral oxycodone / day, 8 mg oral hydromorphone / day, 25 mg oral oxymorphone / day, or an equivalent analgesic dose of another opioid for one week or longer" (fda.gov / downloads / drugs / drugsafety / postmarketdrugsafetyinformationforpatientsandproviders / ucm311290.pdf). Well-established animal models for assessing opioid tolerance exist.

[0031] Given that gelling proteins reduce opioid tolerance, some details and results of the experimental studies presented in this article have been explained. For example, although not intended to be restrictive, Figure 1 Figure 2 illustrates the interpretation of data from conducted morphine tolerance studies. While not wishing to be bound by any particular theory, Figure 1 The diagram illustrates the circulatory nature of inflammation triggered by multiple stimuli, where astrocytes and microglia produce microparticles (MPs) that travel through the glial lymphatic system. In this explanation, MPs travel from the perivascular pathway of the central nervous system to the deep cervical lymph nodes and subsequently into the systemic circulation. CNS-derived MPs from astrocytes that express platelet response protein-1 induce neutrophil activation and produce additional MPs, some expressing filamentous actin (F-actin). F-actin MPs lead to autoactivation of neutrophils, exacerbating the inflammatory process, and neutrophils adhering to cerebral microvasculature trigger further astrocyte and microglia activation. F-actin MPs can be disrupted by plasma coagulants (pGSN), and when this occurs, the concentration of pGSN in the bloodstream decreases. Figure 2 provides a non-limiting interpretation of some of the findings presented in this paper. Figure 2A This explanation elucidates some of the research findings and demonstrates that brain-derived melanoma can initiate a cyclical neuroinflammatory process. This explanation is consistent with... Figure 1 The difference lies in the fact that it provides more details about why F-actin MPs can promote inflammation. Studies, including those published here, have shown that F-actin-positive particles contain high concentrations of NOS2 (the inflammatory form of nitric oxide synthase) and interleukin-1β. Plasma coagulants destroy these MPs, and in the process, plasma coagulant levels decrease. Figure 2B A graph showing the reduction in plasma coagulants observed in morphine-tolerant mice and mice injected with F-actin-positive MP obtained from morphine-tolerant mice is provided.

[0032] Determine the risk and / or presence of opioid tolerance.

[0033] This invention partially includes a method for reducing the risk of opioid tolerance in a subject, a non-limiting example of which is morphine tolerance. Certain embodiments of the risk reduction method of this invention include administering a gelling agent to a subject identified as being at risk of developing opioid tolerance, and administering the gelling agent in an amount that effectively reduces the risk of opioid tolerance in the subject. The effectiveness of the method of this invention in reducing the risk in a subject can be determined by comparing the results of administering the gelling agent to the subject with control results. In some embodiments of this invention, administering the gelling agent to a subject reduces the risk of opioid tolerance in the subject compared to a control risk of developing opioid tolerance, wherein the control risk is the risk of developing opioid tolerance in a subject under substantially the same conditions without the administration of the gelling agent. In some embodiments of the method of this invention, determining the effectiveness of the method of this invention in reducing the risk of opioid tolerance in a subject may include observing the characteristics of the subject, observing the effectiveness of pain management with opioids, observing the behavior of the subject, etc.

[0034] In some implementations, the subject at risk of developing opioid tolerance is the subject to be administered opioids. In some implementations, the opioids are administered over a period of time. As a non-limiting example, opioids may be administered to treat a subject over a period of time, during which one or more opioids may be repeatedly administered to the subject over days, weeks, months, or longer. The term "repeatedly" as used herein means more than once. It should be understood that repeated administration to a subject may occur at a regular schedule (one non-limiting example being every 4 to 6 hours), and in some cases, repeated administration may occur at a more irregular time. The amount of opioid, the frequency of opioid administration to the subject, and / or the duration of opioid administration to the subject can lead to opioid tolerance in the subject.

[0035] In some embodiments of the method of the present invention, determining that a subject is at risk of developing opioid tolerance is based at least in part on the current or upcoming opioid regimen administered to the subject. For example, if the subject is currently receiving an opioid regimen, the current dose of opioid administered to the subject, the frequency of opioid administration, and / or the duration of opioid administration can be used to assess the risk or presence of opioid tolerance in the subject. Based at least in part on such risk assessment, the determination that a subject is at risk of developing opioid tolerance, or that a subject has opioid tolerance, some embodiments of the method of the present invention can be used to prevent, reduce, or eliminate opioid tolerance in a subject.

[0036] In cases where a subject is to be treated with an opioid regimen, the dose of opioid to be administered to the subject, the frequency of opioid administration, and / or the duration of opioid administration can be used to assess the risk of opioid tolerance in the subject. Based at least in part on the assessment of the risk of opioid tolerance determined for the subject to be administered the opioid regimen, some embodiments of the method of the present invention can be used to reduce or eliminate opioid tolerance in the subject.

[0037] In one instance, although not intended to be restrictive, a subject currently being treated with a morphine regimen comprising oral administration of morphine at a dose of at least 60 mg / day for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days or longer may be identified as being at risk of developing morphine tolerance, and therefore at risk of having morphine tolerance. In another non-limiting instance, a subject receiving oral administration of morphine at a dose of at least 60 mg / day for the anticipated 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days or longer may be identified as being at risk of developing morphine tolerance, and therefore at risk of having morphine tolerance. In yet another non-limiting instance, a subject receiving oral oxycodone at least 30 mg / day for at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more days may be assessed and identified as being at risk of developing oxycodone tolerance or having oxycodone tolerance.

[0038] Other characteristics of the subject may also be used to determine whether the subject is at risk of developing opioid tolerance or has opioid tolerance. Some non-limiting examples of such characteristics include, but are not limited to: the subject's medical history, a description of the subject's own behavior and drug use, the subject's behavior (one non-limiting example of which is known drug use and / or abuse), blood tests to determine the opioid level in the subject, the presence of adverse reactions in the subject when the subject is given opioids, and the lack of analgesia in the subject after administration of opioids. Using the methods disclosed herein and / or conventional methods known in the art, those skilled in the art related to pain management and healthcare will be able to assess the subject's risk of opioid tolerance and / or the presence of opioid tolerance in the subject.

[0039] In addition to characteristics that can be assessed in a subject, it should be understood that the risk of opioid tolerance in a subject can be determined based on the treatment regimen to be administered to the subject. For example, if a subject is to be given an opioid pain management regimen, the parameters of that regimen can be used to assess whether the subject is at risk of opioid tolerance. Therefore, a subject's current or upcoming opioid treatment can indicate the risk of opioid tolerance.

[0040] Gel protein agent

[0041] As described elsewhere in this article, it has now been established that administration of colloid protein agents to subjects can treat opioid tolerance in subjects. Colloid proteins are highly conserved, multifunctional proteins, initially described in the cytosol of macrophages and subsequently identified in many vertebrate cells [see, for example, Piktel E. et al., Int J Mol Sci 2018; 19:E2516; Silacci P. et al., Cell Mol Life Sci 2004; 61:2614-23]. A unique characteristic of colloid proteins is that their gene expression encodes a spliced ​​variant of a different plasma isoform (pGSN), which is secreted into the extracellular fluid and differs from its cytoplasmic counterpart (cGSN) by expressing an additional 25 amino acid sequence. pGSN typically circulates in mammalian blood at concentrations of 200 to 300 µg / ml, making it one of the most abundant plasma proteins.

[0042] As used herein, the term "gel sol agent" refers to a composition comprising gel sol molecules. In some embodiments of the methods of the present invention, the gel sol molecule may be a functional fragment or functional derivative of a full-length natural parental gel sol molecule. In some embodiments of the present invention, the gel sol agent comprises only one or more of the following: a gel sol molecule, a functional fragment thereof, or a functional derivative of a gel sol molecule. In some embodiments of the present invention, the gel sol agent may comprise one or more additional components, some non-limiting examples of which are detectable labels, carriers, delivery agents, etc. In some aspects of the present invention, the gel sol molecule is plasma gel sol (pGSN), and in some cases, the gel sol molecule is cytoplasmic GSN. The gel sol molecule included in the compositions and methods of the present invention may be a recombinant gel sol molecule.

[0043] As used herein, the term "gel sol agent" refers to a compound comprising an exogenous gel sol molecule. The term "exogenous" as used herein when referring to a gel sol molecule means the gel sol molecule applied to the object, even if the same gel sol molecule, which can be referred to as an endogenous gel sol molecule, is naturally present in the object. The gel sol agent included in the methods of the present invention can be a wild-type gel sol molecule, for example, whose amino acid sequence is listed herein as SEQ ID NO: 1, GenBank accession number X04412, an isotype, analog, functional variant, functional fragment, or functional derivative of the gel sol molecule. It should be understood that in some embodiments of the invention, the applied gel sol molecule is a gel sol polypeptide, and in some embodiments of the methods of the present invention, the applied gel sol molecule is a polynucleotide encoding a gel sol polypeptide.

[0044] Some embodiments of the method of the present invention include the application of a "gelatin analogue," which, as used herein, refers to a compound that is functionally substantially similar to natural gelatin or fragments thereof. Gelatin analogues comprise a biologically active amino acid sequence substantially similar to that of gelatin and may have substituted, deleted, extended, replaced, or otherwise modified sequences that have biological activity substantially similar to that of gelatin. For example, a gelatin analogue is an analogue that does not have the same amino acid sequence as gelatin but is sufficiently homologous to retain the biological activity of gelatin. Biological activity can be determined, for example, by determining the properties of the gelatin analogue and / or by determining the ability of the gelatin analogue to reduce or prevent opioid tolerance in a subject, and in some embodiments, the ability to reduce or prevent morphine tolerance in a subject. Gelatin biological activity assays are known to those skilled in the art.

[0045] Some embodiments of the method of the present invention include fragments of gel sol proteins. The term "fragment" means any portion of a gel sol molecule that includes a segment of gel sol providing at least a portion or substantially all of the biological activity level of the "parent" gel sol protein. The term "gel sol fragment" means a gel sol fragment made from any source, such as naturally occurring peptide sequences, synthetic or chemically synthesized peptide sequences, and genetically modified peptide sequences. The term "parent" as used herein when referring to gel sol fragments or derivative molecules means the gel sol molecule from which the sequence of the fragment or derivative is derived.

[0046] In some embodiments of the method of the present invention, the sol-gel protein fragment is a functional fragment and retains at least some, and up to all, the functions of its parent sol-gel protein molecule. In some embodiments, the method of the present invention may include the application of a “variant” of the sol-gel protein. The sol-gel protein variants used herein may be compounds that are substantially similar in structure and biological activity to the natural sol-gel protein or fragments thereof. In some aspects of the invention, the sol-gel protein variant is referred to as a functional variant and retains at least some, and up to all, the functions of its parent sol-gel protein molecule.

[0047] Gel protein derivatives are also considered for inclusion in some embodiments of the methods of the present invention. A “functional derivative” of a gel protein is a derivative having a biological activity substantially similar to that of the gel protein. “Substantially similar” means activities that are quantitatively different but identical in nature. For example, a functional derivative of a gel protein may contain the same amino acid backbone as the gel protein, but may also contain other modifications such as post-translational modifications, such as bound phospholipids or covalently linked carbohydrates, depending on the necessity of such modifications for the implementation of the therapeutic methods of the present invention. As used herein, the term also means chemical derivatives comprising gel proteins. Such derivatives can improve the solubility, absorption, biological half-life, etc., of gel proteins. The derivatives can also reduce the toxicity of gel proteins, or eliminate or reduce any undesirable side effects of gel proteins, etc. Derivatives capable of mediating such effects, and particularly chemical moieties, are disclosed in Remington, The Science and Practice of Pharmacy, 2012 (edited by Allen, Loyd V., Jr., 22nd edition). Procedures for coupling such moieties to molecules such as gel proteins are well known in the art. The term “functional derivative” is intended to include “fragments,” “variants,” “analogs,” or “chemical derivatives” of gel proteins.

[0048] certain treatment strategies

[0049] Some embodiments of the method of the present invention include administering a gelling agent to a subject suffering from opioid tolerance or identified as being at risk of developing opioid tolerance. In some embodiments of the treatment method of the present invention, the gelling agent comprises a gelling protein molecule, a functional fragment of a gelling protein molecule, or a functional derivative of a gelling protein molecule. In some embodiments of the present invention, the administered gelling agent comprises plasma gelling protein (pGSN). In some embodiments of the present invention, the administered gelling agent may comprise a recombinant gelling protein molecule.

[0050] Some embodiments of the method of the present invention include administering a gelling agent to a subject in an amount that effectively reduces the risk of opioid tolerance in the subject, reduces the severity of opioid tolerance present in the subject, and / or eliminates opioid tolerance present in the subject. In some embodiments, the therapeutically effective amount of the gelling agent refers to an amount of gelling agent administered to the subject sufficient to prevent the onset and / or progression of opioid tolerance in the subject. Compared to the control risk percentage of opioid tolerance, administration of gelling agents reduced the risk of opioid tolerance due to previous, current, or future opioid use by at least 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%, and 43% respectively. %, 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%. For example, if a subject has a 50% risk of developing opioid tolerance due to ongoing opioid administration, administering an effective amount of a gelling agent to the subject could reduce that 50% risk to less than 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or reduce it to 0% risk.

[0051] In some embodiments of the method of the present invention, the therapeutically effective amount of the gelling agent for treating opioid tolerance in a subject is defined as the amount of gelling agent applied to the subject sufficient to reduce or eliminate opioid tolerance present in the subject. Compared to a control level of opioid tolerance, application of the gelling agent can reduce opioid tolerance in subjects with opioid tolerance by at least 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%, and 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%. In some embodiments, the control level is the level in subjects with opioid tolerance who have not received the gelling agent. In some embodiments, the control level for the subjects is the level of opioid tolerance in the subjects before the administration of the gelling agent.

[0052] The timing and amount of gelling agent administered to the subject can be determined based on the subject's medical condition and pain relief needs. In some embodiments, the gelling agent is administered to the subject one or more times before, during, and after opioid administration. The gelling agent can be administered once or multiple times to subjects identified as requiring such treatment. Multiple administrations of the gelling agent mean administering the gelling agent to the subject 2, 3, 4, 5, 6, 7, 8, 9, 10, or more times. It should be understood that the administration of the gelling agent can be combined with additional treatments for the subject, which in some embodiments is opioid administration, and in some embodiments may include additional opioid-reducing treatments and / or additional non-opioid pain relievers.

[0053] Pain relief treatment options

[0054] As described herein, certain embodiments of the invention include selecting a method for pain relief treatment of a subject based at least in part on the presence of opioid tolerance in the subject or the risk of the subject developing opioid tolerance. In some embodiments of the invention, the selected treatment regimen is administered to a subject requiring such treatment and includes administering an effective amount of a gelling agent to treat opioid tolerance in the subject. In some embodiments, the gelling agent is a soluble gelling protein. In some embodiments, the gelling agent is a recombinant gelling protein. In some embodiments of the pain relief treatment methods of the invention, an opioid substance that causes opioid tolerance in the subject, another opioid substance, or a non-opioid pain medication may also be administered to the subject one or more times before, substantially simultaneously with, and after the administration of the gelling agent. As a non-limiting example of an opioid substance in some embodiments of the methods of the invention, the opioid substance is morphine or a morphine derivative, or may be an opioid substance listed elsewhere herein, or a derivative thereof. In some embodiments of the methods of the invention, the administered opioid substance is a pharmaceutical opioid composition. In some embodiments of the method of the present invention, the opioid is administered under the guidance of a healthcare professional. In some embodiments of the method of the present invention, the opioid is administered by the subject or a person who is not a healthcare professional. In some embodiments of the method of the present invention, the opioid is not administered under the guidance of a healthcare professional.

[0055] As a non-limiting example, in some embodiments of the method of the present invention, the opioid substance is morphine. In some embodiments of the method of the present invention, the morphine administered to the subject is a pharmaceutical morphine composition and is administered under the guidance of a healthcare professional. In some embodiments of the method of the present invention, the morphine is administered by the subject himself or by a person who is not a healthcare professional. In some embodiments of the method of the present invention, the morphine is not administered under the guidance of a healthcare professional.

[0056] Some embodiments of the method of the present invention include selecting pain relief treatment for a subject known to have opioid tolerance (also referred to herein as a subject "having" or "has" opioid tolerance). Some embodiments of the method may, but must not, include determining which one or more opioids are the basis of the subject's opioid tolerance. As used herein in relation to a subject's opioid tolerance, the term "basis" means one or more opioids administered to the subject that result in the presence of opioid tolerance. In some embodiments, a healthcare professional or other person administers one or more opioids that are the basis of the subject's opioid tolerance. In some embodiments, the subject may self-administer one or more opioids that are the basis of the subject's opioid tolerance.

[0057] Some embodiments of the pain relief treatment selection method of the present invention include (1) selecting a gelling agent treatment regimen for a subject, wherein the gelling agent treatment regimen includes administering a composition comprising a gelling agent in an amount effective in treating opioid tolerance in the subject, and (2) selecting a pain relief treatment regimen for a subject, wherein the pain relief regimen includes administering one or more opioids and / or different pain relievers to the subject. In some cases, the method of selecting a pain relief treatment regimen further includes determining the identity of one or more opioids as the basis for the subject's opioid tolerance, and optionally selecting one or more opioids and / or different pain relievers based at least in part on the identified one or more opioids as the basis for the subject's opioid tolerance.

[0058] Some aspects of the present invention include selecting a pain relief treatment for a subject at risk of developing opioid tolerance. Such a selection method includes: (1) selecting a gelling agent treatment regimen for the subject, wherein the gelling agent treatment regimen includes administering a composition comprising a gelling agent in an amount that effectively reduces the risk of opioid tolerance in the subject; and (2) selecting a pain relief treatment regimen for the subject, wherein the pain relief treatment regimen includes administering one or more opioids and / or different pain relievers to the subject.

[0059] In some embodiments of the method of the present invention, selecting a pain relief treatment regimen further includes administering the selected pain relief treatment regimen 2, 3, 4, 5, 6, 7, 8 or more times to the subject, and administering the selected gelling agent treatment regimen 2, 3, 4, 5, 6, 7, 8 or more times to the subject. In some embodiments of the method of the present invention, administering the gelling agent treatment regimen and the selected pain relief treatment regimen may include administering the gelling agent before administering one or more opioids and / or different pain relievers to the subject. Some embodiments of the method of the present invention include administering the gelling agent substantially simultaneously with administering one or more opioids and / or different pain relievers to the subject. Some embodiments of the method of the present invention include administering the gelling agent at a time after administering one or more opioids and / or different pain relievers to the subject. It should be understood that, in some embodiments of the method of the present invention, the pain relief treatment regimen may include multiple administrations of a gelling agent to the subject and / or multiple administrations of one or more opioids and / or other pain relievers, and the timing may include administration of the gelling agent at one or more times before, substantially simultaneously with, and after the administration of one or more opioids and / or different pain relievers.

[0060] In some alternative methods of the present invention, the coagulant agent comprises coagulant molecules, and optionally, the coagulant molecules are recombinant coagulant molecules and may be plasma coagulant protein (pGSN).

[0061] Compared to the control level of opioid tolerance, administration of the selected gellingin treatment regimen and the selected pain relief treatment regimen reduced opioid tolerance in subjects. In some cases, the control level is the presence, absence, and / or severity of opioid tolerance in subjects who received the selected pain relief treatment regimen but not the selected gellingin treatment regimen. Compared to opioid tolerance in controls who received the selected pain relief treatment regimen but not the selected gellingin treatment regimen, administration of the selected pain relief treatment reduced opioid tolerance in subjects by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. As described elsewhere in this document, methods known in the art can be used to determine opioid tolerance, such as, but not limited to, identifying or measuring one or more opioid tolerance symptoms in a subject and / or determining the severity of opioid tolerance in a subject.

[0062] Applying the selected pain-relieving treatment of the present invention reduces the pain of the subject, which may also be referred to herein as improving the subject's pain relief. In some embodiments of the pain-relieving treatment method of the present invention, compared with the pain-relieving efficacy of the selected pain-relieving treatment in a control subject who received the selected pain-relieving treatment but did not receive the selected gellingin treatment, applying the gellingin agent with the selected gellingin treatment increases the pain-relieving efficacy of the selected pain-relieving treatment in the subject by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, 200% or more.

[0063] Therapeutic compositions and methods and monitoring efficacy

[0064] The method of the present invention includes generating a therapeutic effect in subjects suffering from or at risk of developing opioid tolerance to reduce and treat opioid tolerance. The term "therapeutic effect" as used herein when referring to an agent (e.g., a gelling agent) means the therapeutic effect of the gelling agent when administered to a subject suffering from or at risk of developing opioid tolerance. The therapeutic effect of the gelling agent (also referred to herein as a "response" to the treatment method of the present invention) can be determined, for example, by detecting one or more physiological effects of the treatment, such as a reduction or disappearance of physiological characteristics or symptoms of opioid tolerance after administration of the treatment. Further means of monitoring and assessing opioid tolerance in subjects, and ways of assessing and determining one or more of the level, severity, changes in severity, etc., of opioid tolerance in subjects, are known in the art. Some non-limiting examples of physiological symptoms of opioid tolerance include, but are not limited to, decreased sensitivity to opioid pain relief treatments, respiratory depression, behavioral characteristics, etc., which are known in the art and routinely used in the assessment of opioid tolerance. Some non-limiting examples of ways to identify and / or monitor opioid use in subjects (the presence of opioids can indicate opioid tolerance in subjects) are tests such as blood tests to determine the level of opioids in subjects, physical assessments of subjects, and signs of opioid intoxication in subjects (e.g., respiratory depression, disorientation, etc.).

[0065] In a non-limiting instance, it can be determined that subjects receiving chronic opioid administration for pain control (especially at high doses) require higher amounts or doses of opioids to relieve pain compared to other subjects not receiving such administration. It can be assumed that subjects with opioid tolerance have a lower tolerance for opioid-induced respiratory depression than for analgesia. This means that the different amounts or doses of opioids required to achieve substantially equivalent levels of pain relief in subjects could result in higher levels of respiratory depression in opioid-tolerant subjects compared to non-opioid-tolerant subjects. Therefore, patients tolerant to opioids may have an increased risk of respiratory depression when receiving sufficient amounts or doses of opioids to achieve substantially the same level of pain relief as those tolerant to opioids and those not tolerant to opioids (see, for example, CJ Hayhurst, & ME Durieux, Anesthesiology February 2016, Vol. 124, 483–488.).

[0066] Some embodiments of the method of the present invention may further include determining the efficacy of the administered treatment regimen. For example, the level of opioid tolerance in the subject may be determined, and changes therein may be monitored. In some embodiments, determining the level of opioid tolerance includes behavioral assessment. In some embodiments, the level of opioid tolerance in the subject is determined, and this level indicates the regression and / or reduction in the severity of opioid tolerance in the subject. When used herein as reference to opioid tolerance, the term "severity" means the level of opioid tolerance in the subject. As a non-limiting example, the severity of opioid tolerance in a subject who has been given opioids for one week may be less than in subjects who have been given opioids for a longer period of time (e.g., 2, 3, 4, 5, 6, or more weeks).

[0067] In some embodiments of the invention, the subject's opioid tolerance is assessed at one or more of the following times: before the administration of the gelling agent, during the administration of the gelling agent, and after the administration of the gelling agent. Such an assessment can be used to determine the efficacy of the gelling agent in the subject.

[0068] In some embodiments, the method of the present invention includes administering an effective amount of a gelling agent to a subject suffering from or at risk of opioid tolerance, said effective amount producing a therapeutic effect to reduce the severity of opioid tolerance in the subject. For subjects identified as suffering from or at risk of opioid tolerance, the gelling agent may be administered in combination with other treatments selected in their treatment regimen.

[0069] The methods and compositions of the present invention can be used to treat opioid tolerance. As used herein, when used in connection with opioid tolerance, the term "treatment" and variations thereof can refer to preventive treatment that reduces the likelihood or risk of a subject developing opioid tolerance, and / or can refer to treatment after a subject has developed opioid tolerance. In some embodiments of the methods of the present invention, the treatment method reduces the risk of a subject developing opioid tolerance. In some embodiments, the treatment method of the present invention can be used to eliminate or improve existing opioid tolerance in a subject, prevent existing opioid tolerance in a subject from becoming more severe, and / or slow the progression of opioid tolerance in a subject compared to the progression of opioid tolerance in the absence of the treatment method of the present invention.

[0070] This invention partially includes pharmaceutical compositions comprising a gelling agent and one or more opioids. The gelling agent may be a gelling protein molecule, which may be a recombinant gelling protein molecule. In some embodiments, the gelling protein molecule is plasma gelling protein (pGSN). In some embodiments, the opioid is morphine, heroin, hydromorphone, hydroxymorphone, fentanyl, methadone, oxycodone, hydrocodone, codeine, buprenorphine, dextromethorphan, dextropropoxyphene, loperamide, meperidine, opioid, carfentanil / carfentanil, morphone, tapentadone, tramadol, or meperidine. In some embodiments, the pharmaceutical composition comprising the gelling agent may also comprise an opioid, so that the gelling agent and the opioid can be administered together to the subject. It should be understood that administration of the gelling agent and the opioid can be carried out in a separate pharmaceutical composition.

[0071] Objects and Samples

[0072] As used herein, the object can be a vertebrate, including but not limited to humans, mice, rats, guinea pigs, rabbits, cattle, dogs, cats, horses, goats, and non-human primates such as monkeys. The object can be a mammal. In some embodiments, the object is any human or non-human recipient of the gelling agent or pharmaceutical composition comprising a gelling agent described herein. In some embodiments, the object is any human or non-human recipient of the gelling agent or pharmaceutical composition comprising a gelling agent described herein. In some embodiments, the object is any human or non-human recipient of the gelling agent therapeutic regimen and pain relief treatment regimen of the present invention.

[0073] In some aspects of this invention, the object can be a domesticated animal, a wild animal, or an agricultural animal. Therefore, this invention can be used to treat opioid tolerance in both human and non-human subjects. For example, the methods and compositions of this invention can be used in veterinary applications and human treatment protocols. In some embodiments of this invention, the object is a human. In some embodiments of this invention, the object suffers from or is at risk of developing opioid tolerance, and the object requires treatment with the methods of this invention. In some embodiments of the methods of this invention, the object is an animal model of opioid tolerance.

[0074] In some embodiments, the subject treated with one embodiment of the method of the present invention is an adult subject administering opioids to treat pain, a non-limiting example of which is moderate or severe pain. In some embodiments, the subject treated with one embodiment of the method of the present invention is an elderly subject. In some embodiments, the elderly subject does not require treatment for frailty. In some embodiments, the subject treated with one embodiment of the method of the present invention is a subject abusing one or more opioids. In some embodiments, the subject treated with one embodiment of the method of the present invention does not require treatment for an infection with gelosin. In some embodiments, the subject treated with one embodiment of the method of the present invention does not have an infection. In some embodiments, the subject treated with one embodiment of the method of the present invention does not have a decompression-related disease or condition. In some embodiments, the subject treated with one embodiment of the method of the present invention does not have multiple sclerosis.

[0075] Assessment and comparison

[0076] The opioid tolerance characteristics of subjects can be compared with control values ​​of opioid tolerance characteristics. Control values ​​can be predetermined values, which can take various forms. They can be a single cutoff value, such as the median or mean. They can be established based on comparison groups, such as a group of individuals with opioid tolerance, a group of individuals who have received treatment for opioid tolerance, a group of individuals who have not yet received treatment for opioid tolerance, etc. Another example of a comparison group could be a group of subjects with one or more symptoms or diagnoses of opioid tolerance, and a group of subjects without one or more symptoms or diagnoses of opioid tolerance. Of course, the predetermined values ​​will depend on the specific group selected. Therefore, the selected predetermined values ​​can take into account the category to which the individuals belong. Those skilled in the art can select the appropriate category using only routine experiments.

[0077] In the method of the present invention, controls can be used to compare characteristics of different control groups, characteristics of the subject, and characteristics of the control group, etc. Comparisons can be made between the subject and the control, or between one control and another, based on relative differences. For example, but not intended to be limiting, physiological symptoms in subjects treated with the gelling agent of the treatment method of the present invention can be compared with physiological symptoms in a control group that has not received the gelling agent. This comparison can be expressed in relative terms; for example, if insensitivity to the administered opioid is a characteristic of opioid tolerance, the determination of opioid sensitivity in subjects treated with the treatment method of the present invention, including the administration of the gelling agent, can be compared with the control level of opioid sensitivity in subjects that have not received the gelling agent. In some embodiments, a suitable control is a subject not treated with the treatment method of the present invention. The comparison between the treated subject and the control may include comparing the opioid tolerance level between the treated subject and a selected control. In some cases, it can be determined that the opioid tolerance level of the subject treated with the method of the present invention is lower than that of a selected control, wherein the comparison shows that the opioid tolerance level in the subject is reduced by as much as 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%, or 42% compared to the control. 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%.

[0078] It should be understood that, in addition to predetermined values, controls can also be material samples tested in parallel with the experimental material. Some examples include samples from a control group or control samples prepared for testing in parallel with the experimental sample; and controls can be samples from subjects before, during, or after treatment with an embodiment of the method or composition of the present invention. Thus, one or more characteristics identified for subjects at risk of developing opioid tolerance or who have opioid tolerance can be used as “control” values ​​for those characteristics in the subject at later times.

[0079] Timing of application

[0080] Some embodiments of the present invention include pretreatment of subjects who do not yet have opioid tolerance at the time of treatment. In some embodiments, the pretreatment of the subject occurs at some point before the subject receives an opioid treatment regimen assessed to place the subject at risk of developing opioid tolerance. Some embodiments of the treatment methods of the present invention include administering an effective amount of a gelling agent to a subject at risk of developing opioid tolerance, wherein the gelling agent is administered just before the administration of the opioid treatment regimen, or for a maximum of 1, 2, 3, 4, 5, 6, 12, 18, 24, 48, 72, 96, 120, 144 hours or more before the administration of the opioid to the subject. In some embodiments, the gelling agent is administered to the subject at the same time as the opioid is administered to the subject. In some embodiments, the gelling agent is administered after the administration of the opioid to the subject. In some embodiments, the subject receives the gelling agent of the treatment method of the present invention at two or three of the times before, during, and after the administration of the opioid to the subject. It should be understood that subjects identified as currently receiving or about to receive opioid treatment and who are likely to develop opioid tolerance at some point may be given gelling agents as a preventative treatment to reduce the likelihood of opioid tolerance development in the subjects.

[0081] Pharmaceutical Compositions and Delivery

[0082] Some embodiments of the method of the present invention include a method for treating opioid tolerance in an individual (hereinafter interchangeably referred to as the subject) requiring such treatment, and the treatment includes administering a therapeutically effective amount of a gellingin agent to the subject. In one non-limiting example, a gellingin (also referred to herein as a gellingin agent) is administered to a subject requiring treatment for opioid tolerance. In some embodiments, the gellingin is administered intravenously or using another suitable method of administration, some non-limiting examples of which are listed elsewhere herein. A representative example of a gellingin form is recombinant gellingin. Some embodiments of the method of the present invention include a method for prophylactic treatment of an individual susceptible to opioid tolerance, comprising the step of administering a therapeutically effective amount of a gellingin agent to the subject.

[0083] Gel sols may be administered to a subject as a pharmacological composition (also referred to herein as a pharmaceutical composition), meaning that the gel sol is administered as a sterile composition that may contain one or more additional components. The gel sol may be administered in an effective amount that produces the desired response at a weight or volume unit suitable for administration to the subject. Opioids may be administered to a subject as a pharmacological composition (also referred to herein as a pharmaceutical composition), meaning that the composition is sterile and may contain one or more additional components. Opioids may be administered in an effective amount that relieves pain in the subject. The dosage of gel sols and opioids administered to the subject can be selected based on various parameters, particularly the method of administration used and the condition of the subject. Other factors include the desired duration of treatment. In cases where the response in the subject is insufficient upon administration of the initial dose, a higher dose may be used within the limits of the subject's tolerance (or an effective higher dose via a different, more localized route of delivery). The dosage of the medication may be adjusted by an individual physician, healthcare practitioner, or veterinarian; particularly in the event of any complications. The therapeutically effective dose of gelling agents typically ranges from 0.01 mg / kg to about 1000 mg / kg, from about 0.1 mg / kg to about 200 mg / kg, from about 0.2 mg / kg to about 20 mg / kg, from about 0.5 mg / kg to about 10 mg / kg, or from about 1.0 mg / kg to about 5 mg / kg, administered in one or more daily doses for one or more days. In some embodiments of the invention, the therapeutically effective dose ranges from 1 mg / kg to 100 mg. The effective dose of opioids for pain relief can be determined using routine procedures known to those performing healthcare and / or veterinary care.

[0084] The method of the present invention includes administering a gelling agent and / or an opioid to a subject. Methods of administering the pharmaceutical composition are known in the art, and the methods disclosed herein, as well as others known in the art, can be used in the method of the present invention to efficiently deliver the pharmaceutical composition and / or agent of the present invention to desired tissues, cells, or body fluids. The manner of administration and dosage can be adjusted by an individual physician, healthcare practitioner, or veterinarian, particularly in the event of any complications. The absolute amount administered will depend on a variety of factors, including the choice of the material used for administration, whether the administration is a single or multiple dose, and individual subject parameters, including the subject's age, physical condition, body type, weight, severity of opioid tolerance in the subject, existing, ongoing, and / or planned opioid regimens, etc. These factors can be determined and addressed using only routine experiments.

[0085] Various administration methods known to those skilled in the art can be used to effectively deliver the pharmaceutical compositions of the present invention comprising a gelling agent and / or comprising one or more opioid substances to a subject to produce a desired therapeutic effect. Methods of administration of the pharmaceutical compositions of the present invention may include oral, sublingual, oral, intranasal, intravitreal, intracavitary, intravenous, intramuscular, intrathecal, intraperitoneal, subcutaneous, percutaneous, intradermal, topical, rectal, vaginal, intrasynovial, intraocular, or inhalation administration. The present invention is not limited to the specific administration methods disclosed herein. Standard references in the art (e.g., Remington, The Science and Practice of Pharmacy, 2020, edited by Adeboye Adejare, 23rd edition) provide administration methods and formulations for delivering various pharmaceutical preparations and formulations in a drug carrier. Other administration methods known in the art may be used in the methods of the present invention to administer one or more pharmaceutical compositions to a subject. In some embodiments of the methods of the present invention, administration methods in which the dosage, administration schedule, administration site, manner of administration, etc., may differ from those shown herein.

[0086] Administration of one or more pharmacological compositions of the present invention to mammals other than humans (e.g., for testing or research purposes or for veterinary therapeutic purposes) may be carried out under substantially the same conditions as described herein. It should be understood that the present invention is applicable to both humans and animal subjects. Therefore, the present invention is intended for use in animal husbandry and veterinary medicine, as well as in human treatment.

[0087] When administered, the pharmaceutical compositions of the present invention are applied in a pharmaceutically acceptable amount and as pharmaceutically acceptable compositions. The term "pharmaceutical acceptable" means a non-toxic material that does not interfere with the effectiveness of the bioactivity of the active ingredient. Such formulations may typically contain one or more salts, buffers, preservatives, compatible carriers, aqueous solutions, water, etc., and optionally other therapeutic agents. Pharmaceutically acceptable carriers may contain one or more diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials known in the art. As used herein, the term "pharmaceutical acceptable" means a molecular entity and composition that does not produce adverse, allergic, or other adverse reactions when properly administered to animals (e.g., humans). The preparation of pharmaceutical compositions comprising gelling agents, opioids, and / or other medicaments will be known to those skilled in the art according to this disclosure, as illustrated in Remington: The Science and Practice of Pharmacy, 23rd edition, Academic Press, 2020.

[0088] Exemplary pharmaceutically acceptable carriers are known and conventionally used by those skilled in the art. In some embodiments of the invention, such formulations may comprise salts; when used in pharmaceuticals, the salts may be pharmaceutically acceptable, but non-pharmaceutical salts can be readily used to prepare their pharmaceutically acceptable salts and are not excluded from the scope of the invention. Such pharmacologically and pharmaceutically acceptable salts include, but are not limited to, those prepared from acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, citric acid, formic acid, malonic acid, succinic acid, etc. Furthermore, pharmaceutically acceptable salts may be prepared as alkali metal or alkaline earth metal salts, such as sodium, potassium, or calcium salts. The pharmaceutical compositions of the invention may also optionally contain suitable preservatives, such as benzalkonium chloride, chlorobutanol, p-hydroxybenzoate, and thimerosal.

[0089] If desired, pharmaceutical preparations or pharmaceutical compositions may be combined with pharmaceutically acceptable carriers. As used herein, the term "pharmaceutically acceptable carrier" means one or more compatible solid or liquid fillers, diluents, or encapsulating substances suitable for administration to humans. The term "carrier" refers to a natural or synthetic organic or inorganic component combined with an active ingredient to facilitate application. Pharmaceutically acceptable carriers that may be included in the pharmaceutical compositions of the present invention include diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials known in the art. Exemplary pharmaceutically acceptable carriers are described in U.S. Patent No. 5,211,657, and other pharmaceutically acceptable carriers are known to those skilled in the art. In some embodiments of the invention, such preparations may comprise salts, buffers, preservatives, compatible carriers, aqueous solutions, water, etc. When used in pharmaceuticals, salts may be pharmaceutically acceptable, but non-pharmaceutical salts may be readily used to prepare their pharmaceutically acceptable forms and are not excluded from the scope of the invention. Components of the pharmaceutical composition may also be miscible with the pharmaceutical preparations of the present invention and with each other in a manner that does not significantly impair the desired pharmaceutical efficacy. The pharmaceutical compositions used in the methods of the present invention may contain suitable buffers, as described above, including but not limited to: acetates, phosphates, citrates, glycine, borates, carbonates, bicarbonates, hydroxides (and other bases) and pharmaceutically acceptable salts of the aforementioned compounds.

[0090] The pharmaceutical compositions used in the methods of this invention are readily available in unit dosage forms and can be prepared by any method known in the pharmaceutical field. All methods include the step of associating the active agent with a carrier constituting one or more excipients. Generally, the composition is prepared by homogeneously and tightly associating the active compound with a liquid carrier, a finely fragmented solid carrier, or both, and subsequently shaping the product (if desired).

[0091] Compositions suitable for oral administration may exist as discrete units, such as capsules, tablets, pills, or lozenges, each containing a predetermined amount of an active compound (e.g., a gelling agent). Other compositions include suspensions in aqueous or non-aqueous liquids, such as syrups, elixirs, emulsions, or gels.

[0092] Orally administered pharmaceutical formulations can be obtained in the form of solid excipients, optionally by grinding the resulting mixture, and, if desired, by processing the granular mixture after adding suitable excipients to obtain tablets or sugar-coated pellet cores. Suitable excipients are particularly fillers, such as sugars including lactose, sucrose, mannitol, or sorbitol; cellulose formulations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP). If desired, disintegrants, such as cross-linked polyvinylpyrrolidone, agar, or alginate or its salts, such as sodium alginate, may be added. Optionally, oral formulations may also be formulated in saline or buffers (i.e., EDTA for neutralizing internal acidic conditions) or may be administered without any carrier.

[0093] Oral dosage forms of one or more of the aforementioned components are also considered. One or more components can be chemically modified to make the oral delivery of the derivative effective. Typically, the chemical modifications considered involve the attachment of at least one moiety to the component molecule itself, wherein said moiety allows (a) inhibition of proteolytic activity; and (b) uptake from the stomach or intestine into the bloodstream. It is also desirable to improve the overall stability of one or more components and increase their circulation time in vivo. Some examples of such moiety include: polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, and polyproline. (Abuchowski and Davis, 1981, “Soluble Polymer-Enzyme Adducts” In: Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley-Interscience, New York, NY, pp. 367–383; Newmark et al., 1982, J. Appl. Biochem. 4:185–189.) Other polymers that can be used are poly-1,3-dioxolane and poly-1,3,6-tioxocane.

[0094] For the pharmaceutical preparation, the site of release can be the stomach, small intestine (duodenum, jejunum, or ileum), or large intestine. Those skilled in the art will obtain formulations that do not dissolve in the stomach but release substances into the duodenum or other parts of the intestine. Preferably, the release will avoid the harmful effects of the gastric environment by protecting the gelling agent or by releasing the biologically active substance outside the gastric environment (e.g., in the intestine).

[0095] Microspheres formulated for oral administration may also be used. Such microspheres are well defined in the art. All formulations intended for oral administration should be at doses suitable for such administration.

[0096] For oral administration, the composition may be in the form of tablets or lozenges formulated in a conventional manner.

[0097] For administration by inhalation, the compounds used according to the invention can be conveniently delivered in the form of an aerosol from a pressurized package or nebulizer using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas). In the case of pressurized aerosols, the dosage unit can be determined by providing a valve for delivering a measured amount. Capsules and cartridges, such as gelatin, for use with inhalers or blowpipes can be formulated into a powder mixture containing the compound with a suitable powder matrix (e.g., lactose or starch).

[0098] This document also considers the delivery of gelling proteins or opioids via the lungs in the methods of the present invention. Gelling proteins and / or opioids are delivered to the lungs of mammals upon inhalation and cross the epithelial lining into the bloodstream.

[0099] Nasal (or intranasal) delivery of the pharmaceutical compositions of the present invention is also considered. Nasal delivery allows the pharmaceutical compositions of the present invention to travel directly into the bloodstream after the therapeutic product is applied to the nose, without requiring the product to deposit in the lungs. Formulations for nasal delivery include those containing dextran or cyclodextran.

[0100] When systemic delivery of the compound is desired, it can be formulated for parenteral administration via injection (e.g., by bolus injection or continuous infusion). Injectable formulations can be presented in unit dose form with added preservatives, for example, in ampoules or multi-dose containers. The composition can be in the form of a suspension, solution, or emulsion in an oily or aqueous carrier and may contain a formulatory agent, such as a suspending agent, stabilizer, and / or dispersant.

[0101] Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compound in a water-soluble form. Alternatively, suspensions of the active compound can be prepared as suitable oily injectable suspensions. Suitable lipophilic solvents or carriers include fatty oils, such as sesame oil; or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injectable suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or reagents that increase the solubility of the compound to allow for the preparation of highly concentrated solutions. Alternatively, the active compound may be in powder form for use prior to preparation with a suitable carrier (e.g., sterile, pyrogen-free water).

[0102] Drugs, including but not limited to sol-gel agents and / or opioids, may be provided within the particles. As used herein, "particles" refers to nanoparticles or microparticles (or in some cases larger) that may consist wholly or partially of the sol-gel agents described herein. The particles may contain the drug within a core surrounded by a coating (including but not limited to enteric coating). The drug may also be dispersed throughout the particle. The drug may also be adsorbed into the particle. The particles may exhibit release kinetics of any order, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof. In addition to drugs, the particles may contain any of those materials conventionally used in the pharmaceutical and medical fields, including but not limited to erodible, non-erodible, biodegradable, or non-biodegradable materials, or combinations thereof. The particles may be microcapsules containing sol-gel proteins in solution or semi-solid form. The particles can be virtually any shape.

[0103] Both non-biodegradable and biodegradable polymeric materials can be used to manufacture particles for delivering pharmaceutical agents, such as gelling agents or opioids. Such polymers can be natural or synthetic. The polymer is selected based on the desired release time period. Of particular interest are bioadhesive polymers, including biodegradable hydrogels described in HS Sawhney, CP Pathak and JA Hubell in Macromolecules, (1993) 26:581-587 (whose teachings are incorporated herein by reference). These include polyhyaluronic acid, casein, gelatin, glutinin, polyanhydride, polyacrylic acid, alginate, chitosan, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(laurate methacrylate), poly(phenyl methacrylate), poly(methyl methacrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).

[0104] The pharmaceutical preparation may be included in a controlled-release system. The term "controlled release" is intended to refer to any formulation containing a drug in which the manner and profile of drug release from the formulation are controlled. This refers to immediately released formulations and non-immediately released formulations, where non-immediately released formulations include, but are not limited to, sustained-release and delayed-release formulations. The term "sustainable release" (also known as "extended release") is used in its conventional sense to refer to a pharmaceutical preparation that provides a gradual release of the drug over an extended period of time, and preferably, but not necessarily, results in a substantially constant blood level of the drug over the extended period of time. The term "delayed release" is used in its conventional sense to refer to a pharmaceutical preparation in which there is a time delay between formulation administration and the release of the drug therefrom. "Delayed release" may or may not involve the gradual release of the drug over an extended period of time, and therefore may or may not be "sustainable release."

[0105] The use of long-term continuous-release implants is particularly suitable for the treatment of chronic conditions. As used herein, “long-term” release means that the implant is constructed and positioned to deliver a therapeutic level of medication for at least 7 days, and preferably 30 to 60 days. Long-term continuous-release implants are well known to those skilled in the art and include some of the aforementioned release systems.

[0106] The methods and compositions of this invention are of significant importance for subject treatment and the clinical development of new treatments. It is also anticipated that clinical researchers will now use the methods of this invention to determine recruitment criteria for human subjects in clinical trials. Healthcare practitioners will select treatment options based on the expected net benefit for the subject. Net benefit is derived from the risk-benefit ratio.

[0107] The dosage of treatment can be varied, for example, by increasing or decreasing the amount of gelling agent applied to the subject, by changing the therapeutic composition, by changing the route of administration, by changing the time of administration, and so on. The effective dosage will vary depending on the specific disease or condition being treated, the age and physical condition of the subject being treated, the severity of the disease or condition, the duration of treatment, the specific route of administration, and similar factors are all within the knowledge and professional skills of the healthcare practitioner.

[0108] Effective dose

[0109] As used herein, the term "effective amount" refers to the treatment methods or compositions of the present invention. The methods of the present invention involve administering a gelling agent and / or an opioid in an amount that is an effective amount. When administered to a subject according to the methods of the present invention, the effective amount of the gelling agent produces a therapeutic effect against opioid tolerance and / or reduces opioid tolerance in the subject. When administered to a subject according to the methods of the present invention, the effective amounts of the gelling agent and the opioid substance produce a therapeutic effect against opioid tolerance and / or reduce pain in the subject.

[0110] An effective amount refers to a dose of gellingin and / or opioid sufficient to provide a medically desired outcome. Some non-limiting examples of agents used in certain embodiments of the compositions and methods of the present invention include, but are not limited to, gellingin agents. It should be understood that the agents of the present invention are intended to treat or prevent opioid tolerance conditions, i.e., they can be used preventively on subjects at risk of developing opioid tolerance. Therefore, an effective amount of a gellingin agent can be an amount that reduces the risk of developing opioid tolerance, slows down, or may completely prevent the occurrence of opioid tolerance. It should be recognized that when the agent is used in acute situations, it is used to prevent one or more medically undesirable outcomes that typically arise from such adverse events.

[0111] The factors involved in determining the effective amount of gelling agents and / or opioids are known to those skilled in the art and can be resolved through routine experiments, including but not limited to clinical trials for dose evaluation. Generally, the maximum dose of the pharmaceutical agent of the present invention (alone or in combination with other therapeutic agents), i.e., the highest safe dose based on reasonable medical judgment, is preferred. However, those skilled in the art will understand that patients may adhere to lower or tolerable doses for medical, psychological, or virtually any other reason.

[0112] The therapeutically effective amount of the medicament of the present invention is an amount that effectively treats an impairment (e.g., opioid tolerance). In the case of opioid tolerance, the desired response is to inhibit the progression of opioid tolerance and / or reduce the severity of opioid tolerance. This may involve only temporarily slowing the progression of opioid tolerance, but it may also include permanently stopping the progression of opioid tolerance. Inhibiting the progression of opioid tolerance and / or reducing the severity of opioid tolerance in a subject may also involve reducing or decreasing the severity of that one or more characteristics of opioid tolerance in the subject by at least 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more compared to a baseline level of the severity of one or more characteristics of opioid tolerance in the subject. The efficacy of the administered amount can be monitored by conventional diagnostic methods known to those skilled in the art. The expected response to opioid tolerance treatment can also be to delay or even prevent the onset of opioid tolerance in the subject.

[0113] The method of the present invention includes administering a gelling agent in an effective amount for treating opioid tolerance. An effective amount is a dose of gelling agent sufficient to provide a medically desired outcome. The therapeutically effective amount of the pharmacological composition of the present invention is an amount that effectively treats a condition such as opioid tolerance. In the case of opioid tolerance, the desired response may be to suppress the onset and / or progression of opioid tolerance, and / or to reduce the severity of opioid tolerance, and / or to reduce the level of opioid tolerance. This may involve only temporarily slowing the progression of opioid tolerance, but it may also include stopping the progression of opioid tolerance. The efficacy of the gelling agent in treating opioid tolerance in subjects can be monitored using conventional diagnostic methods known to those skilled in the art. The desired response to treatment of opioid tolerance may also be prevention of the onset of opioid tolerance.

[0114] medicine box

[0115] The invention also contemplates the use of a pillbox. In some aspects of the invention, the pillbox may contain one or more vials for pharmaceutical preparations, vials for pharmaceutical preparation diluents, gelling agents, and opioids. Vials containing a diluent for pharmaceutical preparations are optional. The diluent vials may contain a diluent, such as physiological saline, for diluting a concentrated solution or lyophilized powder of a gelling agent. Instructions may include instructions for mixing a specific amount of diluent with a specific amount of concentrated pharmaceutical preparation, thereby preparing a final preparation for administration (e.g., but not limited to injection or infusion). Instructions may include instructions for treating a subject with an effective amount of gelling agent. It should also be understood that the container containing the preparation, whether it is a bottle, a vial with a septum, an ampoule with a septum, an infusion bag, etc., may contain markings, such as conventional markings that change color when the preparation has been autoclaved or otherwise sterilized.

[0116] The invention is further illustrated by the following examples, which should not be construed as further limitations. The entire contents of all references cited throughout this application (including bibliographic references, granted patents, published patent applications, and co-pending patent applications) are expressly incorporated herein by reference.

[0117] The following examples are provided to illustrate specific instances of the practice of the invention, but are not intended to limit the scope of the invention. It will be apparent to those skilled in the art that the invention will be applied to a variety of compositions and methods.

[0118] Example

[0119] Example 1

[0120] The study used a withdrawal test protocol (Figure 3A), in which morphine tolerance was induced in mice by administering morphine injections three times daily, followed by a tolerance test on day four. Tolerance was measured as the length of time it took for a mouse to remove its tail from warm water (a negative stimulus). Figure 3B The results obtained using this morphine tolerance animal model are shown. Results Figure ( Figure 3B The figure shows the time required for mice to be removed from warm water by their tails, with measurements taken at baseline (without morphine injection) and subsequently after cumulative morphine doses of 0.5, 1, 2, and 3 mg / kg IP.

[0121] The results showed that the control mice exhibited a slower response to having their tails removed because they were injected with any dose of morphine (0.5 to 3 mg / kg), while the morphine-tolerant mice responded to warm water by having their tails removed, even after being injected with morphine (0.5, 1, or 2 mg / kg).

[0122] Example 2

[0123] The study aimed to assess the possible relationship between morphine tolerance microparticles and CSF flow in the brain. Figure 4 Schematic diagrams and charts are provided. The top diagram shows the direction of CSF flow in the brain, the bottom left diagram shows the lymphatic system in the mouse brain, and the bottom right diagram shows a chart of particle concentrations determined in controls and morphine-tolerant subjects. Figure 4 The glial lymphatic system and the conventional lymphatic system are shown. Fluid from both systems flows to the deep cervical lymph nodes. A study was conducted measuring microparticles (MPs) in the deep cervical lymph nodes. Results showed that MPs were significantly elevated in morphine-tolerant mice (in...). Figure 4 (As shown in the middle, bottom right).

[0124] Further studies were conducted to evaluate particulate matter and morphine tolerance. Some results were found in... Figure 5 The diagram shows the number of microparticles present in the deep cervical lymph nodes of mice when they were in a morphine-tolerant state and when they underwent a tolerance regimen (see Figure 3). Additionally, several drugs antagonizing different inflammatory pathways were selected and injected into mice. Procedures included injections of: PEGylation polymer B (0.7 μl of a 0.3% solution [w / v] IV / g [PEGylation polymer B is an agent that destroys most MPs when injected into the bloodstream], anaphylactin (100 mg / kg IP [anaphylactin is a drug that blocks interleukin-1β receptors], methylnaltrexone (1 mg / kg IP [methylnaltrexone is an opioid receptor blocker that does not cross the blood-brain barrier], or neutropenic animals [mice were treated with antibodies to selectively remove polymorphonuclear leukocytes (also known as neutrophils) from the bloodstream].

[0125] Example 3

[0126] The study aimed to examine whether antinociceptive tolerance (as shown in Figure 3) could be prevented by treating mice with agents that inhibit particle elevation (e.g., ...). Figure 5 (As shown). Figure 6 The results shown represent the time required for mice to be removed from warm water by their tails, measured at baseline and subsequently after cumulative morphine doses of 0.5, 1, 2, and 3 mg / kg IP. Studies were conducted after mice received either sterile saline daily (control) or 20 mg / kg morphine for two consecutive days followed by 50 mg / kg morphine on the third day. Figure 6 As shown, mice were also given anaerobic agonist (100 mg / kg IP), methylnaltrexone (1 mg / kg IP), or PEG-telopolymer B (0.7 μl of 0.3% solution [w / v] IV / g). Data are mean ± SE of repeated measures, p < 0.05, two-way ANOVA. The results showed that MP levels were significantly higher in morphine-tolerant mice compared with control mice, morphine-tolerant mice treated with PEG-telopolymer B, anaerobic agonist, methylnaltrexone, and morphine-tolerant mice with neutropenia.

[0127] Example 4

[0128] Studies were conducted to evaluate the presence of the microparticles at elevated levels in morphine-tolerant mice (see Example 3). Figure 7 Western blot analysis of the precursor and mature forms of IL-1β is shown. Plasma samples containing 100,000 MP were centrifuged at 21,000 × g for 1 hour. The precipitate was retained for Western blot analysis, and the supernatant was centrifuged at 100,000 g for 1 hour so that the precipitate could also be Western blotted to assess the relative concentrations of precursor and mature IL-1β. The data below the figure show the IL-1β concentrations (pg / 1 million MP) from whole plasma and the supernatant and precipitate fraction after 21,000 g centrifugation. The results shown in the table indicate that IL-1β is elevated in tolerant mice, and almost all IL-1β is present within the microparticles.

[0129] Example 5

[0130] To assess blood-borne methadone levels in opioid-tolerant subjects. Such studies were conducted in which blood samples were collected from patients who had started methadone treatment for opioid use disorder and from age-matched controls who were not taking the medication. Figure 8The results showed that, compared with control levels, blood-derived MP levels were elevated in opioid-tolerant patients who entered methadone treatment.

[0131] Example 6

[0132] A study was conducted to assess glial lymphatic flow. Figures 9A to 9C provide photographs and graphs illustrating measurements of brain glial lymphatic flow assessed by gadolinium (Gd)-contrast MRI. Figure 9A highlights the signal near the Galenian vein and the deep cervical lymph nodes. Flow was quantified using 1 mm target regions at these sites, and the signal was normalized by including a 60 mM Gd vial located beneath the mouse skull within the scanned field of view. Images of control and tolerant mice are shown in... Figure 9B Figure 98C shows a graph of quantified glial lymphatic flow measured in 5 mice. The vertical axis represents the normalized Gd signal, and the horizontal axis represents the time following Gd injection in the mice. The MRI machine was programmed to acquire images every 5 minutes and subsequently report the values. Therefore, the horizontal axis is shown at consecutive 5-minute intervals. The data indicate that mice in morphine tolerance exhibited increased glial lymphatic flow compared to control mice that received only sterile saline injections.

[0133] Example 7

[0134] The study aimed to evaluate the effects of injecting MP obtained from morphine-tolerant mice into naïve mice. Figure 10 The results of an experiment conducted by injecting F-actin-positive MP from morphine-tolerant mice into naïve mice are shown. Overall, the results indicate that morphine injection increases glial lymphatic flow, and this contributes to the increase in MP present in the deep cervical lymph nodes and blood. The results of the PEG-tunomer B injection study suggest that blood-derived MP is essential for morphine tolerance. Further research indicates that, in particular, F-actin-positive MP triggers persistent inflammation. Figure 10 This indicates that when mice were injected with F-actin MP from tolerant mice, they showed somewhat similar increases in glial lymphatic flow (left panel) and also showed neuroinflammation (right Western blot).

[0135] Example 8

[0136] Subjects are assessed and, at least in part, based on the opioid regimen to be administered to them, their risk of opioid tolerance is determined. In addition to administering opioids, subjects are given an effective amount of a gelling agent to reduce or eliminate opioid tolerance episodes. In some studies, the opioids administered include morphine, heroin, hydromorphone, hydroxymorphone, fentanyl, methadone, oxycodone and hydrocodone, codeine, buprenorphine, dextromethorphan, dextropropoxyphene, loperamide, meperidine, opioids, carfentanil / carfentanil, morphone, tramadol, tapentadol and / or meperidine. A treatment regimen including the administration of a gelling agent is selected to treat opioid tolerance in subjects, at least in part, based on the therapeutic opioid regimen to be administered, and both the opioid treatment regimen and the gelling agent treatment regimen are administered to the subjects. Subjects were evaluated before, during, and after the opioid and gelling agent regimens, and it was determined that administration of the gelling agent reduced and / or prevented opioid tolerance in the subjects. As a result of the gelling agent regimen, subjects showed reduced opioid tolerance compared to control opioid tolerance.

[0137] Example 9

[0138] Subjects are assessed and, at least in part, based on the morphine regimen to be administered to them, their risk of morphine tolerance is determined. In addition to administering morphine, an effective amount of a gelling agent is administered to the subject to reduce or eliminate episodes of morphine tolerance. Based at least in part on the therapeutic morphine regimen administered to the subject, a treatment regimen including the administration of a gelling agent is selected to treat morphine tolerance. The morphine treatment regimen and the gelling agent treatment regimen are administered to the subject.

[0139] Subjects were evaluated before, during, and after the morphine and gelosin regimens, and it was determined that administration of the gelosin regimen treated morphine tolerance in the subjects. As a result of the gelosin regimen, subjects showed reduced morphine tolerance compared to control morphine tolerance.

[0140] Subjects were evaluated before, during, and after the morphine and gelling agent regimens, and administration of the gelling agent reduced and / or prevented opioid tolerance in the subjects. As a result of the gelling agent regimen, subjects showed reduced opioid tolerance compared to control opioid tolerance.

[0141] Example 10

[0142] Subjects were identified as having opioid tolerance, and an effective amount of gelling agent was administered to them as treatment for opioid tolerance. The gelling agent was administered to subjects in an amount that effectively reduced or eliminated opioid tolerance. In some studies, opioid tolerance was one or more of the following: oxycodone tolerance, fentanyl tolerance, morphine tolerance, hydromorphone tolerance, methadone tolerance, hydrocodone tolerance, tramadol tolerance, morphine tolerance, heroin tolerance, codeine tolerance, buprenorphine tolerance, dextromethorphan tolerance, dextropropoxyphene tolerance, loperamide tolerance, meperidine tolerance, opioid tolerance, carfentanil / carfentanil tolerance, morphine tolerance, tapentadol tolerance, and meperidine tolerance. A treatment regimen including the administration of a gelling agent was selected for subjects to treat opioid tolerance. The gelling agent treatment regimen was administered to the subjects. Compared with control opioid tolerance, the application of gelling agents was effective in reducing and / or preventing opioid tolerance in subjects.

[0143] Example 11

[0144] Subjects were identified as having morphine tolerance and were administered an effective amount of gelling agent as treatment for morphine tolerance. Gelling agent was administered to subjects in an amount that effectively reduced or eliminated morphine tolerance. A treatment regimen including gelling agent administration was selected for the subjects to treat morphine tolerance. The gelling agent treatment regimen was administered to the subjects. Compared with control opioid tolerance, administration of gelling agent was effective in reducing and / or preventing morphine tolerance in subjects.

[0145] Equivalent scheme

[0146] While several embodiments of the invention have been described and illustrated herein, those skilled in the art will readily conceive of a variety of other ways and / or structures for performing the functions described herein and / or obtaining the results and / or one or more advantages described herein, and each such variation and / or modification is considered to be within the scope of the invention. More generally, those skilled in the art will readily understand that all parameters, dimensions, materials, and configurations described herein are intended to be exemplary, and actual parameters, dimensions, materials, and / or configurations will depend on one or more specific applications using the teachings of this invention. Those skilled in the art will recognize, or can determine, many equivalents of the specific embodiments of the invention described herein using only conventional experimentation. Therefore, it should be understood that the foregoing embodiments are presented by way of example only, and that the invention may be practiced in ways other than those specifically described and claimed within the scope of the appended claims and their equivalents. The invention relates to each individual feature, system, article, material, and / or method described herein. Furthermore, any combination of two or more of such features, systems, articles, materials, and / or methods is included within the scope of the invention if they do not contradict each other.

[0147] All definitions defined and used herein should be understood to take precedence over dictionary definitions, definitions in referenced literature, and / or the usual meaning of the defined terms.

[0148] As used herein, when the term "comprising / including / contains" is used in conjunction with the claims and / or description, a noun without a quantifier may mean "one / a," but it is also consistent with the meaning of "one / a or more / a," "at least one / a," and "one / a or more than one / a." Some embodiments of the invention may consist of or substantially consist of one or more elements, method steps, and / or methods of the invention. It is contemplated that any method described herein can be implemented in relation to any other method described herein.

[0149] As used herein, unless explicitly stated otherwise, the term "or" in the claims is used to mean "and / or," but this disclosure supports the definition of "and / or" referring only to alternatives. The phrase "and / or" as used herein in the specification and claims should be understood to mean "any one or both" of the elements so connected, i.e., elements that coexist in some cases and exist separately in others. Unless explicitly stated otherwise, additional elements may optionally exist besides those specifically indicated by the "and / or" clause, whether related to or unrelated to those specifically indicated.

[0150] As used herein, “comprising / including / containing” and its variations shall be understood to imply inclusion of the stated item, element, or step, or a group of items, elements, or steps, but not to exclude any other item, element, or step, or a group of items, elements, or steps, unless the context requires otherwise. Similarly, “another” or “other” may mean at least a second or more identical or different elements of the claims or components thereof.

[0151] All references, patents and patent applications, and publications cited or referenced in this application are incorporated herein by reference in their entirety.

Claims

1. A method for treating opioid tolerance in subjects, comprising: A composition comprising a gelling agent is administered to subjects requiring such treatment in an amount that is effective in treating opioid tolerance.

2. The method of claim 1, wherein the opioid tolerance is morphine tolerance.

3. The method of claim 1 or 2, wherein the gelling protein agent comprises a gelling protein molecule, a functional fragment thereof, or a functional derivative thereof.

4. The method of claim 3, wherein the coagulant molecule is plasma coagulant protein (pGSN).

5. The method of claim 3, wherein the gel sol protein molecule is a recombinant gel sol protein molecule.

6. The method of claim 1, wherein the composition comprising the gelling protein agent is administered orally, sublingually, intranasally, intravenously, intramuscularly, intrathecally, intraperitoneally, subcutaneously, intradermally, superficially, rectally, vaginally, intrasynovially, or intraocularly, or by inhalation.

7. The method of claim 1, wherein administration of the gelling agent has a greater therapeutic effect on opioid tolerance in the subject compared with the control treatment effect against opioid tolerance.

8. The method of claim 7, wherein the control treatment effect is equivalent to the effect on opioid tolerance in the subject in the absence of administration of the gelling agent.

9. The method of claim 1, wherein applying the gelling agent reduces opioid tolerance in the subject by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to the severity of opioid tolerance in a control without the application of the gelling agent.

10. The method of claim 1, wherein the object is a mammal.

11. The method of claim 10, wherein the mammal is a human.

12. The method of claim 1, wherein the gelling agent is applied to the object 1, 2, 3, 4, 5, 6, 7, 8 or more times.

13. The method of claim 1, wherein applying the gelling agent increases the analgesic efficacy of the opioid applied to the subject by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to the analgesic efficacy of a control without the application of the gelling agent.

14. The method of claim 1, further comprising administering an opioid to a subject to which the composition comprising the gelling agent is applied.

15. The method of claim 1, wherein the applied gelling agent reduces the level of opioid tolerance in the subject.

16. The method of claim 1, wherein the applied gelling agent prevents opioid tolerance in the subject.

17. The method of claim 1, wherein the opioid substance comprises: Oxycodone, fentanyl, morphine, opium, hydromorphone, methadone, or hydrocodone.

18. A method for selecting pain relief treatment for subjects with opioid tolerance, the method comprising: (a) Selecting a gelling agent treatment regimen for the subject, comprising administering a composition comprising a gelling agent at an amount effective in treating opioid tolerance in the subject; (b) Selecting a pain relief treatment regimen for the subject, which includes administering one or more opioids and / or different pain relievers to the subject.

19. The method of claim 18, further comprising determining the identity of the opioid, and optionally selecting the one or more opioids and / or the different pain relievers based at least in part on the identity of the opioid.

20. The method of claim 18, further comprising applying the selected pain relief treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8 or more times.

21. The method of claim 18, further comprising applying the selected gelling protein treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8 or more times.

22. The method of claim 18, wherein the administration of the gelling agent treatment regimen and the pain relief treatment regimen comprises administering the gelling agent before, simultaneously with, and after administering the one or more opioids and / or the different pain relievers to the subject.

23. The method of claim 18, wherein the coagulant agent comprises coagulant molecules, wherein the coagulant molecules are optionally recombinant coagulant molecules and optionally plasma coagulant protein (pGSN).

24. The method of claim 18, wherein the administration of the selected gellingin treatment regimen and the selected pain relief treatment regimen have a greater therapeutic effect against opioid tolerance in the subjects compared with the control treatment effect against opioid tolerance, optionally, wherein the control treatment effect is the effect against opioid tolerance in subjects who received the selected pain relief treatment regimen and did not receive the selected gellingin treatment regimen.

25. The method of claim 18, wherein the application of the gelling agent reduces opioid tolerance in the subject by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to a control opioid tolerance in the absence of the gelling agent.

26. The method of claim 25, wherein the opioid tolerance is measured based on one or more of the following: the presence of one or more opioid tolerance symptoms in the subject and the severity of opioid tolerance in the subject.

27. The method of claim 18, wherein the object is a mammal, optionally a human.

28. The method of claim 18, wherein applying the gelling agent increases the pain-relieving efficacy of the selected pain-relieving treatment regimen in the subjects by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, 200%, or more compared to the pain-relieving efficacy of a control without the application of the gelling agent.

29. The method of claim 18, wherein the opioid comprises: Oxycodone, fentanyl, morphine, hydromorphone, methadone, or hydrocodone.

30. A method for selecting pain relief treatment for a subject at risk of developing opioid tolerance, the method comprising: (a) Selecting a gelling agent treatment regimen for the subject, comprising administering a composition comprising a gelling agent in an amount that effectively reduces the risk of opioid tolerance in the subject; (b) Selecting a pain relief treatment regimen for the subject, which includes administering one or more opioids and / or different pain relievers to the subject.

31. The method of claim 30, further comprising applying the selected pain relief treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8 or more times.

32. The method of claim 30, further comprising applying the selected gelling protein treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8 or more times.

33. The method of claim 30, wherein the administration of the gelling agent treatment regimen and the pain relief treatment regimen comprises administering the gelling agent to the subject before, simultaneously with, and after administering one or more of the one or more opioids and / or the different pain relievers.

34. The method of claim 30, wherein the gelling agent comprises gelling protein molecules, wherein the gelling protein molecules are optionally recombinant gelling protein molecules.

35. The method of claim 34, wherein the coagulant molecule is plasma coagulant protein (pGSN).

36. The method of claim 30, wherein administering the gellingin treatment regimen and the pain relief treatment regimen reduces the risk of opioid tolerance in the subjects compared to a control treatment effect against the risk of opioid tolerance, optionally wherein the control treatment effect is equal to the effect against opioid tolerance in subjects who received the pain relief treatment regimen but did not receive the gellingin treatment regimen.

37. The method of claim 30, wherein, compared with opioid tolerance in a control group not treated with the gelling agent, treatment with the gelling agent reduces the risk of opioid tolerance in the subject by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.

38. The method of claim 30, wherein the object is a mammal, optionally a human.

39. The method of claim 30, wherein applying the gelling agent increases the pain-relieving efficacy of the selected pain-relieving treatment regimen in the subjects by at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, 200%, or more compared to the pain-relieving efficacy of a control without the application of the gelling agent.

40. The method of claim 30, wherein the opioid comprises: Oxycodone, fentanyl, morphine, hydromorphone, methadone, or hydrocodone.

41. A pharmaceutical composition comprising a gelling agent and an opioid.

42. The pharmaceutical composition of claim 41, wherein the coagulant agent comprises a coagulant molecule, wherein the coagulant molecule is optionally a recombinant coagulant molecule and optionally a plasma coagulant protein (pGSN).

43. The pharmaceutical composition of claim 41, wherein the opioid is morphine, heroin, hydromorphone, hydroxymorphone, fentanyl, methadone, oxycodone, hydrocodone, codeine, buprenorphine, dextromethorphan, dextropropoxyphene, loperamide, meperidine, opioid, carfentanil / carfentanil, morphone, tapentadol, tramadol, or meperidine.

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