Gelsolin and opioid tolerance

Abstract

The present invention relates to compositions and methods for treating opioid tolerance in a subject, and methods for selecting a pain relief treatment regimen for a subject suffering from opioid tolerance and / or at risk of suffering from opioid tolerance. Embodiments of the therapeutic methods of the invention include administering a gelsolin agent to produce a therapeutic effect in a subject against opioid tolerance.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480070458.6 (22) Application Date 2024.11.08 (30) Priority Data 63 / 547,801 2023.11.08 US (85) PCT International Application Entering National Phase Date 2026.05.06 (86) PCT International Application Application Data PCT / US2024 / 055075 2024.11.08 (87) PCT International Application Publication Data WO2025 / 101865 EN 2025.05.15 (71) Applicant University of Maryland, Baltimore, USA (72) Inventor Stephen R. Thom (74) Patent Agency Beijing J&J Intellectual Property Agency Co., Ltd. 11227 Patent Attorney Wu Xiaojing (51) Int.Cl. A61K 38 / 17 (2006.01) A61K 31 / 485 (2006.01) A61K 45 / 06 (2006.01) A61P 25 / 04 (2006.01) A61P 25 / 36 (2006.01) (54) Invention Title: Gel Protein and Opioid Tolerance (57) Abstract: This invention relates to compositions and methods for treating opioid tolerance in subjects, and 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 the present invention include administering a gel protein agent to produce a therapeutic effect against opioid tolerance in subjects. Claims (3 pages), Description (24 pages), Sequence List (electronic publication), Drawings (10 pages), CN 122138836 A, 2026.06.02, CN 1 22 13 88 36 A. 1. A method for treating opioid tolerance in a subject, comprising: administering a composition comprising a gelling agent to a subject requiring such treatment in an amount 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 agent comprises a gelling protein molecule, a functional fragment thereof, or a functional derivative of a gelling protein molecule. 4. The method of claim 3, wherein the gelling protein molecule is plasma gelling protein (pGSN). 5. The method of claim 3, wherein the gelling protein molecule is a recombinant gelling protein molecule.6. The method of claim 1, wherein the composition comprising the gelling 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 against opioid tolerance in the subject compared to a control treatment effect against opioid tolerance. 8. The method of claim 7, wherein the control treatment effect is equivalent to the effect against opioid tolerance in the subject without administration of the gelling agent. 9. The method of claim 1, wherein 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 the severity of opioid tolerance in a control without application of the gelling agent. 10. The method of claim 1, wherein the subject 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 subject 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 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 analgesic efficacy of the opioid in the subject without application of the gelling agent. 14. The method of claim 1, further comprising administering the opioid to the 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 comprises: oxycodone, fentanyl, morphine, opioid, hydromorphone, hydroxymorphone, methadone, or hydrocodone.18. A method 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 to the subject one or more opioid substances and / or different pain relievers. 19. The method of claim 18, further comprising determining the identity of the opioid substance, and optionally selecting the one or more opioid substances and / or the different pain relievers at least in part based on the identity of the opioid substance. 20. The method of claim 18, further comprising administering 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 administering the selected gellingin treatment regimen 2, 3, 4, 5, 6, 7, 8 or more times to the subject. 22. The method of claim 18, wherein administering the gellingin treatment regimen and the pain relief treatment regimen comprises administering the gellingin 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 gellingin agent comprises a gellingin molecule, wherein the gellingin molecule is optionally a recombinant gellingin molecule, and optionally a plasma gellingin (pGSN). 24. The method of claim 18, wherein administering 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 a control treatment effect against opioid tolerance, optionally wherein the control treatment effect is the effect against opioid tolerance in subjects who have received the selected pain relief treatment regimen but have not received 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 subject is a mammal, optionally a human. 28. The method of claim 18, wherein application of the 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. 29. The method of claim 18, wherein the opioid comprises: oxycodone, fentanyl, morphine, hydromorphone, methadone, or hydrocodone. 30. A method 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 that effectively reduces the risk of opioid tolerance in the subject; and (b) selecting a pain relief treatment regimen for the subject, comprising administering to the subject one or more opioids and / or different pain relievers. 31. The method of claim 30, further comprising administering the selected pain relief treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8, or more times. Claims 2 / 3 Page 3 CN 122138836 A 32. The method of claim 30, further comprising administering the selected gelling agent treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8, or more times. 33. The method of claim 30, wherein administering the coagulant treatment regimen and the pain relief treatment regimen comprises administering the coagulant 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 coagulant agent comprises a coagulant molecule, wherein the coagulant molecule is optionally a recombinant coagulant molecule. 35. The method of claim 34, wherein the coagulant molecule is plasma coagulant protein (pGSN).36. The method of claim 30, wherein administering the gelling agent 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 gelling agent treatment regimen. 37. The method of claim 30, wherein administering the gelling agent reduces the risk of opioid tolerance 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%, or 100% compared to opioid tolerance in a control without the gelling agent. 38. The method of claim 30, wherein the subject 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 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 regimen 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 comprises a coagulant molecule, wherein the coagulant molecule is optionally a recombinant coagulant molecule, and optionally a plasma coagulant molecule (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, tapentadone, tramadol, or meperidine.Claims 3 / 3 Page 4 CN 122138836 A Gel Protein and Opioid Tolerance

[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 Interest

[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 U.S. Office of Naval Research. The government enjoys certain rights in this invention. Technical Field

[0005] In some aspects, this invention relates to compositions and methods for reducing opioid tolerance. Background Art

[0006] Opioid tolerance is recognized as a serious problem in healthcare. There is currently no effective method to reduce and / or eliminate opioid tolerance, and there is a widespread need for such a method as an adjunct treatment for moderate and 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 in need of such treatment in an amount effective in treating opioid tolerance in the subject. In some embodiments, 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, superficially, rectally, vaginally, intrasynovially, or intraocularly. It can be administered orally, sublingually, intravenously, subcutaneously, topically, intrathecally, or by inhalation. In some embodiments, administration of the gelling agent has a greater therapeutic effect against opioid tolerance in subjects compared to the control treatment effect against opioid tolerance. In some embodiments, the control treatment effect is equivalent to the effect against opioid tolerance in subjects without the administration of the gelling agent.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 the severity of opioid tolerance in a control group without gelling agent application. In some embodiments, the subjects are mammals. In some embodiments, the mammals are humans. In some embodiments, the gelling agent is applied to the subjects 1, 2, 3, 4, 5, 6, 7, 8, or more times. In some embodiments, the application of a gelling agent increases the analgesic efficacy of the opioid 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 analgesic efficacy of a control without the application of a gelling agent. In some embodiments, the method further includes administering the opioid to the 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 opioids include: oxycodone, fentanyl, morphine, opium, hydromorphone, 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 includes applying the selected gelling protein 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, 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 that effectively reduces 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 includes 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 includes administering the selected coagulant treatment regimen to the subject 2, 3, 4, 5, 6, 7, 8 or more times. In some embodiments, administering the coagulant treatment regimen and the pain relief treatment regimen includes administering the coagulant agent to the subject before, simultaneously with, and after one or more of the administration of 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, the administration of a gelling agent treatment regimen and a pain relief treatment regimen reduces the risk of opioid tolerance in subjects compared to the control treatment effect against the risk of opioid tolerance. Optionally, the control treatment effect is equal to the effect against opioid tolerance in subjects who received the pain relief treatment regimen but not the gelling agent treatment regimen. In some embodiments, the administration of a gelling 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 the gelling agent. In some embodiments, the subjects are mammals; optionally, the subjects are humans. 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 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 coagulant agent comprises coagulant molecules, wherein the coagulant molecules are optionally recombinant coagulant molecules, and optionally plasma coagulant 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.

[0011] These and other features, aspects, and advantages of embodiments of the present 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, in which:

[0012] Figure 1 is a schematic diagram depicting the interpretation of data from a morphine tolerance study. This figure 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 produce 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] Figures 2A and 2B provide schematic diagrams and graphs. Figure 2A is a schematic diagram illustrating the interpretation of some research findings and showing that brain-derived MPs can initiate a cyclical neuroinflammatory process. This interpretation differs from Figure 1 in that it shows more details about why F-actin MPs exhibit pro-inflammatory activity. 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 provides a graph showing the reduction in plasma coagulants in morphine-tolerant mice and mice injected with F-actin-positive MPs obtained from morphine-tolerant mice.Instructions for Use, Page 3 / 24, CN 122138836 A

[0014] Figures 3A to 3B provide a schematic diagram and list 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 is a graph showing the time required for mice to remove their tails from warm water when measured after cumulative doses of morphine injected at baseline (no morphine injection) and subsequently intraperitoneally (IP) at doses of 0.5, 1, 2, and 3 mg / kg. The results showed that controls exhibited a slower response to remove their tails due to injections of arbitrary doses (0.5 to 3 mg / kg) of morphine, while mice in morphine tolerance responded to warm water by removing their tails despite injections of morphine (0.5, 1, or 2 mg / kg).

[0015] Figure 4 provides schematic diagrams and graphs illustrating the following: the top graph shows the direction of CSF flow in the brain, the lower left graph shows the lymphatic system in the mouse brain, and the lower right graph is a graph of particle concentration 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 were conducted in which particle (MP) was measured in the deep cervical lymph nodes, and the results showed that MP was significantly elevated in mice in morphine tolerance (shown in the graph at the lower right).

[0016] Figure 5 provides a graph showing the number of particles present in the deep cervical lymph nodes of mice in morphine tolerance and in mice undergoing a tolerance regimen (see Figure 3) plus injection of multiple drugs to antagonize different inflammatory pathways. 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 polymorphonuclear leukocytes (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 provides a graph illustrating tolerance antagonism and showing that treatment of mice with an agent that prevents particle elevation (as shown in Figure 3) can prevent anti-nociceptive tolerance (as shown in Figure 1). Figure 6 shows the time required for mice to be removed from warm water after cumulative morphine doses of 0.5, 1, 2, and 3 mg / kg IP, measured at baseline and subsequently after these doses. Studies were conducted after receiving daily sterile saline (control) or morphine at 20 mg / kg for two consecutive days and 50 mg / kg on day 3. It is indicated that mice were also given anastomosis (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 measurements, p < 0.05, two-way ANOVA.

[0018] Figure 7 provides images of Western blots probing the precursor form of IL-1β and the mature protein. 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 contents of precursor and mature IL-1β. The data below the figure show the concentrations of IL-1β from whole plasma and the supernatant and precipitate fractions after 21,000 g centrifugation, in pg / 1,000,000 MP. Data are mean ± SE, n = 6. The results shown in the table indicate that IL-1β is elevated in tolerant mice, and 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 provides a table showing results from a study in which blood samples were collected from patients newly admitted to methadone treatment for opioid use disorder and age-matched untreated controls. Data are presented as mean ± SD, n=40 per group, t-test. Results showed elevated levels of blood-derived MP in patients admitted to methadone treatment compared to control levels.

[0020] Figures 9A to 9C provide photographs and graphs showing measurement results of glial lymphatic flow assessment by gadolinium (Gd) contrast MRI, page 4 / 24, CN 122138836 A. 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 area, and the signal was normalized by including a 60 mM Gd vial under the mouse skull within the scanning field. Images of control and tolerant mice are shown in Figure 9B. Figure 9C shows a graph of the quantitative glial lymph flow measured in 5 mice.The vertical axis represents the normalized Gd signal, and the horizontal axis represents the time following Gd injection in mice. The MRI machine was programmed to collect images for 5 minutes and then report the values. Therefore, the horizontal axis is shown as 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 injection.

[0021] Figure 10 provides a schematic diagram, graph, and Western blot image illustrating an experiment conducted by injecting F-actin-positive MP from morphine-tolerant mice into naïve mice. 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-modulator B injection study suggest that blood-derived MP is required for morphine tolerance. Further studies have shown that F-actin-positive MP, in particular, triggers persistent inflammation. Figure 10 shows that when mice were injected with F-actin MP from tolerant mice, they showed somewhat similar increases in glial lymphatic flow (left panel) and neuroinflammation (right Western blot). The number under each band represents the band density relative to the mean ± SD (n = 3 brains / lane) of β-actin from control samples from a replicate study. Western blots detected IBA-1 (which indicates microglia activation), aquaporin-4 (AQP4, a protein that drives glial lymph flow in astrocytes), myeloperoxidase (MPO, showing evidence of neutrophil adhesion to the cerebral vascular system), and CD36 protein (which is associated with neuroinflammation when elevated).

[0022] Brief description of the sequence

[0023] SEQ ID NO: 1 is the amino acid sequence of human plasma coagulant protein having GenBank® accession No. X04412:

[0024] Detailed description

[0025] The present invention is based in part on the unexpected discovery that coagulant protein can be applied to subjects to treat opioid tolerance in subjects. Some embodiments of the method of the present 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. Some aspects of the invention include selecting pain-relieving treatments for subjects known to have opioid tolerance or at risk of developing opioid tolerance. Some embodiments of the method of the present invention can improve the efficacy of pain management in subjects by administering gelling agents. It has 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 opioids used for chronic pain relief or other pain conditions can lead to reduced efficacy. Opioid tolerance is the physical effect of repeated opioid use and can lead to opioid-induced hyperalgesia, which is 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 be developed by administering illicit and / or legal opioids to a subject.

[0028] A variety of opioids are used for analgesia in healthcare and are illicitly used. The term “opioid” as used herein may refer to natural or synthetic opioids. Some non-limiting examples of opioids that can cause 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] Loss of the therapeutic pain relief benefits of opioids due to ongoing or prior opioid use is termed "opioid tolerance," and may necessitate increasing the amount and dosage of opioids administered to maintain satisfactory levels of pain relief in opioid-tolerant individuals. 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 caused by administration of "at least 60 mg of oral morphine / day, 25 mcg of transdermal fentanyl / hour, 30 mg of oral oxycodone / day, 8 mg of oral hydromorphone / day, 25 mg of oral hydroxymorphone / 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 exist for assessing opioid tolerance.

[0031] Given that gellingins reduce opioid tolerance, some details and results from experimental studies presented herein have been interpreted. For example, Figures 1 and 2, while not intended to be restrictive, depict the interpretation of data from conducted morphine tolerance studies. While not wishing to be bound by a particular theory, the schematic diagram in Figure 1 illustrates the circulatory nature of inflammation triggered by multiple stimuli, in which 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. AF-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 herein. Figure 2A illustrates the interpretation of some research findings and shows that brain-derived MPs can initiate a cyclical neuroinflammatory process. This interpretation differs from Figure 1 in that it provides more detail on why F-actin MPs can promote inflammation. Studies, including those disclosed herein, have shown that F-actin-positive granules contain high concentrations of 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 provides a graph showing the reduction in plasma coagulants observed in morphine-tolerant mice and mice injected with F-actin-positive MPs obtained from morphine-tolerant mice.

[0032] Determining the Risk and / or Presence of Opioid Tolerance

[0033] This invention includes, in part, a method for reducing the risk of opioid tolerance in a subject, a non-limiting example of which is morphine tolerance. Some embodiments of the method of reducing risk 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 result of administering a gelling agent to the subject with a control result. In some embodiments of this invention, administering a 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 opioid tolerance in the subject under substantially the same conditions without the administration of a 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 embodiments, the subject at risk of developing opioid tolerance is the subject to be administered opioids. In some embodiments, the opioids are administered over a period of time. As a non-limiting example, opioids may be administered to treat the 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 the 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 opioids, the frequency of administration of opioids to the subject and / or the duration of administration of opioids to the subject may result in the subject developing opioid tolerance to the administered opioids.

[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 administration regimen to the subject. For example, if a subject is currently receiving an opioid regimen, the current dose of opioids 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 the subject is at risk of opioid tolerance, or that the 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 the case of a subject to be treated with an opioid regimen, the dose of opioid to be administered to the subject, the frequency of opioid administration to the subject, and / or the duration of opioid administration to the subject 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 an opioid regimen, some embodiments of the method of the present invention can be used to reduce or eliminate opioid tolerance in the subject. Specification 7 / 24 pages 11 CN 122138836 A

[0037] In one example, although not intended to be limiting, a subject currently being treated with a morphine treatment regimen may be identified as being at risk of developing or having morphine tolerance, said morphine treatment 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. In another non-limiting example, a subject who will be administered morphine orally for 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days or longer at a dose of at least 60 mg / day may be identified as being at risk of developing morphine tolerance, and therefore at risk of having morphine tolerance. In another non-limiting example, a subject who is administered at least 30 mg of oral oxycodone / 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 having 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 having 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 by the subject), blood tests to determine the opioid level in the subject, the presence of adverse reactions in the subject when the subject is given opioids, the lack of analgesia in the subject after the administration of opioids, etc. 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 the characteristics that can be assessed in the subject, it should be understood that the risk of opioid tolerance in the subject can be determined based on the treatment regimen to be administered to the subject. For example, if the 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, the subject's current or upcoming opioid treatment can indicate the risk of the subject developing opioid tolerance.

[0040] Coagulant Protein Agents

[0041] As described elsewhere herein, coagulant protein agents administered to subjects have been identified as therapeutics for opioid tolerance in subjects. Coagulants 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 coagulants 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 a sequence of an additional 25 amino acids. 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 markers, 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 contained 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" is a compound comprising exogenous gel sol molecules. The term "exogenous" as used herein when referring to gelling protein molecules means the gelling protein molecule applied to the object, even if the same gelling protein molecule that can be referred to as an endogenous gelling protein molecule is naturally present in the object. The gelling protein agent included in the method of the present invention can be a wild-type gelling protein molecule, for example, its amino acid sequence listed herein as SEQ ID NO: 1 on page 8 / 24 of the specification, GenBank accession number X04412, isotype, analogue, functional variant, functional fragment, or functional derivative of the gelling protein molecule. It should be understood that in some embodiments of the present invention, the applied gelling protein molecule is a gelling protein polypeptide, and in some embodiments of the method of the present invention, the applied gelling protein molecule is a polynucleotide encoding a gelling protein polypeptide.

[0044] Some embodiments of the method of the present invention include the application of a “gel sol analog”, which, as used herein, refers to a compound that is functionally substantially similar to a natural gel sol protein or a fragment thereof. A gel sol analog comprises a biologically active amino acid sequence substantially similar to that of a gel sol protein and may have substituted, deleted, extended, replaced, or otherwise modified sequences that have biological activity substantially similar to that of a gel sol protein. For example, a gel sol analog is an analog that does not have the same amino acid sequence as a gel sol protein but is sufficiently homologous to retain the biological activity of a gel sol protein. Biological activity can be determined, for example, by determining the properties of the gel sol analog and / or by determining the ability of the gel sol analog 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 a gel sol protein molecule. The term "fragment" means any portion of a colloid molecule that provides a colloid segment that maintains at least a portion or substantially all of the biological activity level of the "parent" colloid protein. The term "colloid protein fragment" means a colloid protein 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 colloid protein fragments or derivative molecules means the colloid protein molecule from which the sequence of the fragment or derivative originates.

[0046] In some embodiments of the method of the invention, the colloid protein fragment is a functional fragment and retains at least some, and at most all, the functions of its parent colloid protein molecule. In some embodiments, the method of the invention may include the application of a "variant" of the colloid protein. The colloid protein variant used herein may be a compound that is substantially similar in structure and biological activity to a natural colloid protein or a fragment thereof. In some aspects of the invention, the colloid protein variant is referred to as a functional variant and retains at least some, and at most all, the functions of its parent colloid 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 a 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 a 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 that include gel proteins.Such derivatives can improve the solubility, absorption, and biological half-life of gelling proteins. The derivatives can also reduce the toxicity of gelling proteins or eliminate or reduce any undesirable side effects of gelling proteins. 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 conjugating such moieties to molecules such as gelling proteins are well known in the art. The term "functional derivative" is intended to include "fragments," "variants," "analogs," or "chemical derivatives" of gelling proteins.

[0048] Certain Treatment Strategies

[0049] Certain embodiments of the methods of the present invention include administering a gelling protein 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 applied gelling agent comprises plasma gelling protein (pGSN). In some embodiments of the present invention, the applied gelling agent may comprise a recombinant gelling protein molecule.

[0050] Some embodiments of the method of the present invention include applying 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 applied 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%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, and 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 the risk of developing opioid tolerance due to ongoing opioid administration is 50%, administering an effective amount of the gelling agent can reduce the risk to less than 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or to 0%.

[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 the amount of gelling agent applied to the subject sufficient to reduce or eliminate opioid tolerance present in the subject.Compared to the control group with opioid tolerance, administration of gelling agents reduced 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%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, and 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 of the gelling agent administration and the amount of gelling agent administered to the subject can be determined based on the subject's medical condition and the subject's need for pain relief. In some embodiments, the gelling agent is administered to the subject at one or more times before, during, and after opioid administration. The gelling agent can be administered once or multiple times to subjects identified as needing such treatment. Multiple applications of the gelling agent mean applying the gelling agent to the subject 2, 3, 4, 5, 6, 7, 8, 9, 10 or more times. It should be understood that the application of the gelling agent can be combined with additional treatments for the subject, which in some embodiments is the administration of an opioid to the subject, and in some embodiments may include the administration of additional opioid-reducing treatments and / or additional non-opioid pain relievers to the subject.

[0053] Pain Relief Treatment Selection

[0054] As described herein, certain embodiments of the invention include methods for selecting pain relief treatments for the 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 present invention, as described on pages 10 / 24 of this specification (CN 122138836 A), 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 method of the present invention, an opioid substance, another opioid substance, or a non-opioid pain medication that causes opioid tolerance in the subject 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 method of the present 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 method of the present invention, the administered opioid substance is a pharmaceutical opioid composition. In some embodiments of the method of the present invention, the opioid substance 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 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 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 a pain relief treatment for a subject known to have opioid tolerance (also referred to herein as a subject “having” opioid tolerance or a subject “has” opioid tolerance). Some embodiments of the method may, but must not, include determining which one or more opioids are the basis for the subject’s opioid tolerance. As used herein, the term "base" in relation to opioid tolerance in a subject 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 form the basis of the subject's opioid tolerance. In some embodiments, the subject may self-administer one or more opioids that form 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 pain relief treatment for a subject at risk of developing opioid tolerance. Such selection methods include: (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, the administration of the gelling agent treatment regimen and the selected pain relief treatment regimen may include administering the gelling agent prior to administering one or more opioids and / or different pain relievers to the subject. Some embodiments of the method of the present invention include administering a gelling agent substantially simultaneously with the administration of 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 following the administration of 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 the gelling agent and / or multiple administrations of one or more opioids and / or other pain relievers to the subject, and the timing may include administering the gelling agent at one or more of the following 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 invention, the gelling agent comprises a gelling protein molecule, and optionally, the gelling protein molecule is a recombinant gelling protein molecule, and may be a plasma gelling protein (pGSN).

[0061] Administration of the selected gelling protein treatment regimen and the selected pain relief treatment regimen to subjects reduces opioid tolerance in subjects compared to a control level of opioid tolerance. In some cases, the control treatment level is the presence, absence, and / or severity of opioid tolerance in subjects who received the selected pain relief treatment regimen but did not receive the selected gelling protein treatment regimen. Compared to a control group receiving the selected pain relief treatment but not the selected gelling agent treatment, the application 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 herein, opioid tolerance can be determined using methods known in the art, such as, but not limited to, identifying or measuring one or more opioid tolerance symptoms in subjects and / or determining the severity of opioid tolerance in subjects.

[0062] The application of the selected pain relief treatment of the present invention reduces the pain of subjects, which may also be referred to herein as improving pain relief in subjects. In some embodiments of the pain relief treatment method of the present invention, compared with the pain relief efficacy of the selected pain relief treatment method in control subjects who received the selected pain relief treatment method but did not receive the selected gellingin treatment method, the application of the gellingin agent with the selected gellingin treatment method increased the pain relief efficacy of the selected pain relief treatment method 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.

[0063] Treatment Compositions and Methods and Monitoring Efficacy

[0064] The method of the present invention includes producing a therapeutic effect in subjects suffering from opioid tolerance or at risk of suffering from opioid tolerance to reduce and treat opioid tolerance. When referring to a pharmaceutical agent (such as a gelling agent), the term "therapeutic effect" as used herein means the therapeutic effect of the gelling agent when administered to a subject who has opioid tolerance or is 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 the reduction or disappearance of physiological characteristics or symptoms of opioid tolerance after administration of the treatment. Other means of monitoring and assessing opioid tolerance in subjects, and methods 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 methods of determining and / or monitoring opioid use in subjects (the presence of opioids can indicate opioid tolerance in subjects) include 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 example, 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 who do not receive chronic opioid administration for pain control. 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 than in non-opioid-tolerant subjects. Therefore, patients with opioid tolerance may have an increased risk of respiratory depression when receiving sufficient amounts or doses of opioids to achieve substantially equivalent levels of pain reduction compared to opioid-tolerant and non-opioid-tolerant subjects (see, for example, C. J. 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 in reference to opioid tolerance, the term “severity” means the level of opioid tolerance in a 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 a subject who has 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, opioid tolerance in a subject 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 end of the administration of the gelling agent. Such an assessment can be used to determine the efficacy of administering the gelling agent to the subject.

[0068] In some embodiments, the method of the invention includes administering an effective amount of the gelling agent to a subject who has opioid tolerance or is 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 having opioid tolerance or at risk of developing 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 may refer to preventive treatment that reduces the likelihood or risk of developing opioid tolerance in a subject, and / or may 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 developing opioid tolerance in a subject. In some embodiments, the treatment method of the present invention may be used to eliminate or improve existing opioid tolerance in a subject to prevent existing opioid tolerance in a subject from becoming more severe, and / or to 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] The present invention includes in part 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, tapentadol, tramadol, or meperidine. In some embodiments, the pharmaceutical composition containing the gelling protein agent may also contain an opioid, so the gelling protein agent and the opioid may be administered together to the target.It should be understood that the administration of the gelling agent and the opioid can be performed in the presence of the gelling agent and the opioid in separate pharmaceutical compositions.

[0071] Subjects and Samples

[0072] As used herein, subjects can be vertebrates, including but not limited to humans, mice, rats, guinea pigs, rabbits, cattle, dogs, cats, horses, goats, and non-human primates, such as monkeys. Subjects can be mammals. In some embodiments, subjects are any human or non-human recipients of the gelling agent or pharmaceutical composition containing the gelling agent described herein. In some embodiments, subjects are any human or non-human recipients of the gelling agent or pharmaceutical composition containing the gelling agent described herein. In some embodiments, subjects are any human or non-human recipients of the gelling agent treatment regimen and pain relief treatment regimen of the present invention.

[0073] In some aspects of the invention, subjects can be domesticated animals, wild animals, or agricultural animals. Therefore, the present invention can be used to treat opioid tolerance in human and non-human subjects. For example, the methods and compositions of the present invention can be used in veterinary applications and human treatment regimens. In some embodiments of the invention, the subject is a human being. In some embodiments of the invention, the subject has opioid tolerance or is at risk of developing opioid tolerance, and the subject requires treatment with the method of the invention. In some embodiments of the method of the invention, the subject is an animal model of opioid tolerance.

[0074] In some embodiments, the subject treated with an embodiment of the method of the 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 an embodiment of the method of the invention is an elderly subject. In some embodiments, the elderly subject does not require treatment for frailty. In some embodiments, the subject treated with an embodiment of the method of the invention is a subject abusing one or more opioids. In some embodiments, the subject treated with an embodiment of the method of the invention does not require treatment for infection with gelosomalin. In some embodiments, the subject treated with an embodiment of the method of the invention does not have an infection. In some embodiments, the subject treated with an embodiment of the method of the invention does not have a decompression-related disease or condition. In some embodiments, the subject treated with an embodiment of the method of the invention does not have multiple sclerosis.

[0075] Assessment and Control

[0076] The opioid tolerance characteristics in the subjects can be compared with control values ​​of opioid tolerance characteristics. The control value can be a predetermined value, which can take many forms. It can be a single cutoff value, such as the median or the mean. It can be established based on comparison groups, such as a group of individuals with opioid tolerance, a group of individuals who have been treated for opioid tolerance, a group of individuals who have not yet been treated 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 ​​may take into account the category to which the individual belongs. 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 subjects, characteristics of control groups, etc. Comparisons can be made between subjects and controls, between one control and another, etc., 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 applied 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 application 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 implementation details (pages 14 / 24, CN 122138836 A), a suitable control is a subject not treated with the treatment method of the present invention. The comparison between the treatment subject and the control may include comparing the opioid tolerance levels between the treatment subject and the 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%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%. 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%.

[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 to be tested 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 invention. Thus, one or more characteristics determined 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 a later time.

[0079] Timing of Application

[0080] Some embodiments of the 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 that is 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 opioids in 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 opioids to the subject. In some embodiments, the gelling agent is administered to the subject at the same time as the administration of opioids. In some embodiments, the gelling agent is administered to the subject after the administration of opioids. In some embodiments, the subject receives the gelling agent of the treatment methods of the present invention at two or three times before, during, and after the administration of opioids. It should be understood that subjects identified as currently being treated with or about to be treated with opioids and who are likely to develop opioid tolerance at some point in time may be administered gelling agents as a prophylactic treatment to reduce the likelihood of opioid tolerance developing in the subject.

[0081] Pharmaceutical Composition and Delivery

[0082] Some embodiments of the method of the present invention include methods 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 gelling agent to the individual. In one non-limiting example, a gelling agent (also referred to herein as a gelling agent) is administered to a subject requiring treatment for opioid tolerance. In some embodiments, the gelling agent 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 gelling agent form is recombinant gelling agent.Some embodiments of the method of the present invention include a method for prophylactic treatment of individuals susceptible to opioid tolerance, comprising the step of administering a therapeutically effective amount of gelling protein to the individual.

[0083] The gelling protein agent may be administered to the subject in the form of a pharmacological composition (also referred to herein as a pharmaceutical composition), meaning that the gelling protein agent is administered as a sterile composition that may contain one or more additional components. The gelling protein agent 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 the subject in the form of 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 the gelling protein agent and the opioid administered to the subject may be selected according to different parameters, particularly according to the method of administration used and the condition of the subject. Other factors include the desired duration of treatment. If 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 agent may be adjusted by an individual physician, healthcare practitioner, or veterinarian; particularly in the event of any complications. The therapeutically effective amount of gellingin 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 amount ranges from 1 mg / kg to 100 mg. The effective amount of opioids for pain relief may be determined using routine procedures known to those performing healthcare and / or veterinary care.

[0084] The method of the invention comprises administering gellingin and / or opioids to the subject. Methods of administering the pharmaceutical composition are known in the art, and the methods disclosed herein, as well as other methods known in the art, can be used in the methods of the present invention to effectively deliver the pharmaceutical composition and / or agent of the present invention to desired tissues, cells, or body fluids. The manner of administration and dosage may 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 dose or multiple doses, and individual subject parameters, including the subject's age, physical condition, body size, weight, severity of opioid tolerance in the subject, and existing, ongoing, and / or planned opioid regimens. 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 containing a gelling agent and / or containing one or more opioids to a subject to produce the 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, surface, 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 a variety of pharmaceutical preparations and formulations in a drug carrier. Other administration methods known in the art can 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, administration method, etc., may differ from those shown herein may be used.

[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 above. It should be understood that the present invention is applicable to both human and animal subjects. Therefore, the present invention is intended for use in animal husbandry and veterinary medicine as well as for human treatment.

[0087] When administered, the pharmaceutical compositions of the present invention are applied in a pharmaceutically acceptable amount and as a pharmaceutically acceptable composition. 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 typically contain one or more salts, buffers, preservatives, compatible carriers, aqueous solutions, water, 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 or composition that, when properly administered to animals (e.g., humans), does not produce adverse, allergic, or other adverse reactions.According to this disclosure, the preparation of pharmaceutical compositions comprising gelling agents, opioids, and / or other medicaments will be known to those skilled in the art, 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 as pharmaceuticals, the salts may be pharmaceutically acceptable, but non-pharmaceutical salts may 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 present invention may optionally also contain suitable preservatives, such as benzalkonium chloride, chlorobutanol, para-hydroxybenzoate, and thimerosal.

[0089] If desired, the pharmaceutical preparation or pharmaceutical composition may be combined with a pharmaceutically acceptable carrier. 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 that is 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 present invention, such formulations may contain 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 salts and are not excluded from the scope of the invention. Components of the pharmaceutical composition may also be miscible with the pharmaceutical agents of the invention and with each other in a manner that does not significantly impair the desired pharmaceutical efficacy. Pharmaceutical compositions used in the methods of the 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] Pharmaceutical compositions used in the methods of the invention may be readily available in unit dosage forms and may 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, compositions are prepared by uniformly and tightly associating an active compound with a liquid carrier, a finely granulated 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, lozenges, each containing a predetermined amount of the 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 may be obtained as solid excipients, optionally by grinding the resulting mixture, and, if desired, by processing the granular mixture after adding suitable adjuvants to obtain tablets or sugar-coated cores. Suitable excipients, particularly fillers, include sugars such as 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). Disintegrants, such as cross-linked polyvinylpyrrolidone, agar, or alginate or its salts, such as sodium alginate, may be added if desired. 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 above-described components are also specifically considered. One or more components may be chemically modified to make oral delivery of the derivative effective. Generally, the chemical modifications considered are at least one part linked to the component molecule itself (page 17 / 24, CN 122138836 A), wherein said part allows (a) inhibition of proteolytic degradation; 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 in vivo circulation time. Some examples of such parts 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, N.Y., 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 formulation, 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 in 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 for oral administration should be at doses suitable for such administration.

[0096] For sublingual 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 spray 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 a valve that provides the delivery dose. Capsules and cartridges, such as gelatin, for inhalers or blowpipes can be formulated as powder mixtures containing the compound with a suitable powder matrix (e.g., lactose or starch).

[0098] Delivery of gelling proteins or opioids via the lungs in the methods of the present invention is also contemplated herein. Gelling proteins and / or opioids are delivered to the lungs of mammals upon inhalation and pass through the epithelial lining into the bloodstream.

[0099] Nasal (or intranasal) delivery of the pharmaceutical compositions of the present invention is also contemplated. Nasal delivery allows the pharmaceutical compositions of the present invention to pass 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 by injection (e.g., by bolus injection or continuous infusion). Injectable formulations may be present in unit dose form with added preservatives, for example, in ampoules or multi-dose containers. The composition may 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 comprise aqueous solutions of the active compound in a water-soluble form. Alternatively, suspensions of the active compound may 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 suspensions for injection 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] Pharmaceutical agents may be provided in the particles, specifically including but not limited to sol-gel agents and / or opioids. As used herein, particles refer to nanoparticles or microparticles (or in some cases larger) that may consist wholly or partially of the sol-gel agent described in this specification, page 18 / 24, CN 122138836 A. The particles may contain the pharmaceutical agent in a core surrounded by a coating (including but not limited to enteric coating). The pharmaceutical agent may also be dispersed throughout the particle. The pharmaceutical agent may also be adsorbed into the particle. The particles may have any level of release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof. In addition to pharmaceutical agents, the particles may also 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 gel sol 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 (e.g., gel sol proteins or opioids). Such polymers may be natural or synthetic polymers. The polymer is selected based on the desired release time period. Particularly interesting bioadhesive polymers include biodegradable hydrogels described in H. S. Sawhney, C. P. Pathak and JA Hubell in Macromolecules, (1993) 26:581-587 (whose teachings are incorporated herein). 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 contained 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 both immediately released and non-immediately released formulations, wherein 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 formulation 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 formulation in which there is a time delay between the administration of the formulation 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 sustained-release implants may be particularly suitable for the treatment of chronic conditions. As used herein, “long-term” release means that the implant is constructed and arranged to deliver a therapeutic level of the drug for at least 7 days, and preferably 30 to 60 days. Long-term sustained-release implants are well known to those skilled in the art and include some of the release systems described above.

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

[0107] The amount of treatment can be varied, for example by increasing or decreasing the amount of gelling agent administered to the subject, by changing the therapeutic composition administered, by changing the route of administration, by changing the time of administration, etc. The effective amount 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 within the knowledge and expertise of the healthcare practitioner.

[0108] Effective Amount

[0109] The term “effective amount” as used herein refers to the treatment method or composition of this invention. The methods of this invention involve administering a gelling agent and / or an opioid in an amount that is an effective amount. When administered to a subject in accordance with the methods of this invention, the effective amount of gelling agent produces a therapeutic effect against and / or reduces opioid tolerance in the subject. When administered to a subject using the method of the present invention, effective amounts of the gelling protein and opioids produce a therapeutic effect against opioid tolerance and / or alleviate pain in the subject.

[0110] Effective amount refers to a dose of gelling protein and / or opioids sufficient to provide the medically desired outcome. Some non-limiting examples of pharmaceutical agents used in certain embodiments of the compositions and methods of the present invention include, but are not limited to, gelling protein agents.It should be understood that the medicaments of the present invention are used to treat or prevent opioid tolerance disorders, i.e., they are preventably used on subjects at risk of developing opioid tolerance. Therefore, the effective amount of the gelling 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 medicaments are used in acute situations, they are 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 the gelling agent and / or opioids are known to those skilled in the art and can be resolved simply by routine experiments, including but not limited to clinical trials for evaluating dosage. Generally, the maximum dose of the medicaments of the present invention (alone or in combination with other therapeutic agents) is preferred, i.e., the highest safe dose based on reasonable medical judgment. 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 medicaments of the present invention is an amount that effectively treats an obstacle (e.g., opioid tolerance). In cases of opioid tolerance, the desired response is to suppress the progression of opioid tolerance and / or reduce its severity. This may involve only temporarily slowing the progression of opioid tolerance, but it may also include permanently stopping its progression. Suppressing the progression of opioid tolerance and / or reducing its severity in a subject may also involve reducing the severity of 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 dose can be monitored using conventional diagnostic methods known to those skilled in the art. The desired response to treatment for opioid tolerance may also be to delay or even prevent the onset of opioid tolerance in a 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 opioid tolerance treatment may also be prevention of opioid tolerance onset.

[0114] Kit

[0115] The invention also contemplates the use of a kit. In some aspects of the invention, the kit may comprise one or more pharmaceutical preparation vials, pharmaceutical preparation diluent vials, the gelling agent, and the opioid. The vial containing the pharmaceutical preparation diluent is optional. The diluent vial may contain a diluent, such as physiological saline, for diluting a concentrated solution or lyophilized powder of the 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 formulation for administration (e.g., but not limited to injection or infusion). Instructions may include instructions for treating subjects with an effective amount of the gelling agent. It should also be understood that containers containing formulations, whether they are bottles, vials with diaphragms, ampoules with diaphragms, infusion bags, etc., may contain markings, such as conventional markings that change color when the formulation has been autoclaved or otherwise sterilized. Specification 20 / 24 pages 24 CN 122138836 A

[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 document references, authorized 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 and 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] Examples

[0119] Example 1

[0120] The study was conducted using a withdrawal test protocol (Figure 3A), in which morphine tolerance was induced in mice by administering three injections of morphine to each mouse 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 shows the results obtained using this morphine tolerance animal model. The results graph (Figure 3B) shows the time required for mice to remove their tails from warm water, where measurements were 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 controls exhibited a slower response to removing their tails because they were injected with any dose of morphine (0.5 to 3 mg / kg), while the morphine-tolerant mice responded to the warm water by removing their tails despite being injected with morphine (0.5, 1, or 2 mg / kg).

[0122] Example 2

[0123] A study was conducted to assess the possible relationship between morphine tolerance microparticles and CSF flow in the brain.Figure 4 provides a schematic diagram and graphs, with the direction of CSF flow in the brain shown in the upper figure, the lymphatic system in the mouse brain shown in the lower left figure, and a graph showing the particle concentrations determined in controls and morphine-tolerant subjects in the lower right figure. Figure 4 shows the glial lymphatic system and the conventional lymphatic system. Fluid from both systems flows to the deep cervical lymph nodes. Studies were conducted in which particle (MP) in the deep cervical lymph nodes were measured. The results showed that MP was significantly elevated in mice in a morphine-tolerant state (shown in Figure 4, lower right).

[0124] Further studies were conducted to evaluate particle and morphine tolerance. Some results are shown in Figure 5, which provides a graph showing the number of particles present in the deep cervical lymph nodes when mice were in a morphine-tolerant state and when subjected to a tolerance protocol (see Figure 3). In addition, a variety of drugs antagonizing different inflammatory pathways were selected and injected into mice. The procedure 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], anaproleukin (100 mg / kg IP) [anaproleukin is an agent 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] Studies were conducted to examine whether antinociceptive tolerance (as shown in Figure 3) could be prevented by treating mice with agents that inhibit microparticle elevation (as shown in Figure 5). The results shown in Figure 6 represent the time required for mice to be removed from warm water by the tail, which was measured at baseline and subsequently after cumulative morphine doses of 0.5, 1, 2, and 3 mg / kg IP. The study was conducted in mice after daily administration of sterile saline (control) or after administration of 20 mg / kg morphine for two consecutive days followed by administration of 50 mg / kg morphine on the third day. As shown in Figure 6, mice were also administered anaspirin (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 elevated in morphine-tolerant mice compared with control, morphine-tolerant mice treated with PEG-telopolymer B, anaspirin, methylnaltrexone, and morphine-tolerant mice with neutropenia.

[0127] Example 4

[0128] Studies were conducted to evaluate the presence of microparticles at elevated levels in morphine-tolerant mice (see Example 3).Figure 7 shows Western blots for probing the precursor form and mature protein of IL-1β. Plasma samples containing 100,000 MP were centrifuged at 21,000 × g for 1 hour. The precipitate was left 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 content of precursor IL-1β and mature IL-1β. The data below the figure show the IL-1β concentration (pg / 1 million MP) from whole plasma and the supernatant and precipitate fraction after centrifugation at 21,000 g. 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] Evaluation of blood-borne MP levels in opioid-tolerant subjects. Such a study was conducted in which blood samples were collected from patients who had started methadone treatment due to opioid use disorder and from age-matched controls who had not taken the drug. Figure 8 shows the results, which indicate that the blood-derived MP level was elevated in opioid-tolerant patients who entered methadone treatment compared with control levels.

[0131] Example 6

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

[0133] Example 7

[0134] Studies were conducted to evaluate the effects of injecting MP derived from morphine-tolerant mice into naïve mice. Figure 10 shows the results of experiments conducted by injecting F-actin-positive MP from morphine-tolerant mice into naïve mice. Overall, the results indicated that morphine injection increased glial lymphatic flow, and this contributed to the increase in MP present in the deep cervical lymph nodes and blood. The PEG-modulator B injection study results suggest that blood-derived MP is essential for morphine tolerance. Further studies showed that, in particular, F-actin-positive MP triggered persistent inflammation.Figure 10 shows 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 were evaluated and, at least in part, determined to be at risk of opioid tolerance based on the opioid regimen to be administered to the subjects. In addition to administering opioids to the subjects, effective amounts of a gelling agent were administered to the subjects to reduce or eliminate the onset of opioid tolerance in the subjects. In some studies, the opioids administered were morphine, heroin, hydromorphone, hydroxymorphone, fentanyl, methadone, oxycodone and hydrocodone, codeine, buprenorphine, dextromethorphan, dextropropoxyphene, loperamide, meperidine, opioids, carfentanil / carfentanil, morphone, tramadol, tapentadol and / or meperidine. Based at least in part on the therapeutic opioid regimen administered to the subject, a treatment regimen including the administration of a gelling agent was selected for the subject to treat opioid tolerance, and the opioid treatment regimen (pages 22 / 24, CN 122138836 A) and the gelling agent treatment regimen were administered to the subject. The subject was evaluated before, during, and after the opioid and gelling agent regimens, and it was determined that the administration of the gelling agent to the subject reduced and / or prevented opioid tolerance in the subject. As a result of the gelling agent regimen, the subject's opioid tolerance was reduced compared to control opioid tolerance.

[0137] Example 9

[0138] The subject was evaluated and, based at least in part on the morphine regimen to be administered to the subject, it was determined that the subject was at risk of morphine tolerance. In addition to administering morphine to the subject, an effective amount of gelling agent was administered to the subject to reduce or eliminate the onset of morphine tolerance in the subject. At least in part, based on the therapeutic morphine regimen administered to the subjects, a treatment regimen including the administration of a gelling agent was selected to treat morphine tolerance in the subjects. The morphine treatment regimen and the gelling agent treatment regimen were administered to the subjects.

[0139] The subjects were evaluated before, during, and after the morphine and gelling agent regimens, and it was determined that the administration of the gelling agent to the subjects treated morphine tolerance. As a result of the gelling agent regimen, the subjects' morphine tolerance was reduced compared to control morphine tolerance.

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

[0141] Example 10

[0142] A subject was identified as having opioid tolerance, and an effective amount of gelling agent was administered to the subject as a treatment for opioid tolerance. The gelling agent was administered to the subject in an amount that would effectively reduce or eliminate opioid tolerance. In some studies, opioid tolerance is one or more of the following: oxycodone tolerance, fentanyl tolerance, morphine tolerance, hydromorphone tolerance, hydroxymorphone 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 the subject to treat opioid tolerance. The gelling agent treatment regimen was administered to the subject. 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 given an effective amount of gelling agent as treatment for morphine tolerance. Gelling agent was applied to subjects in an amount that effectively reduced or eliminated morphine tolerance in subjects. A treatment regimen including the application of gelling agent was selected for the subjects to treat morphine tolerance in subjects. The gelling agent treatment regimen was applied to the subjects. Compared with control opioid tolerance, the application of gelling agent was effective in reducing and / or preventing morphine tolerance in subjects.

[0145] Equivalents

[0146] Although several embodiments of the invention have been described and illustrated herein, those skilled in the art will readily conceive of various 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 experiments. 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 different from the specific description and claims within the scope of the appended claims and their equivalents. This 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 such features, systems, articles, materials, and / or methods do not contradict each other.

[0147] All definitions as defined and used herein should be understood to take precedence over dictionary definitions, definitions in referenced documents, and / or the usual meaning of 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 may be practiced in relation to any other method described herein.

[0149] As used herein, the term “or / or” in the claims is used to mean “and / or” unless explicitly stated otherwise, 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, elements other than those specifically indicated by the “and / or” clause may optionally exist, whether related to or unrelated to those specifically indicated.

[0150] As used herein, “comprising / including / containing” and its variations will be understood to imply inclusion of the stated item, element, or step, or a group of items, elements, or steps, but does not 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 claim elements 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.Instruction manual 24 / 24 pages 28 CN 122138836 A Figure 1 Figure 2A Instruction manual illustration 1 / 10 pages 29 CN 122138836 A Figure 2B Instruction manual illustration 2 / 10 pages 30 CN 122138836 A Instruction manual illustration 3 / 10 pages 31 CN 122138836 A Figure 4 Instruction manual illustration 4 / 10 pages 32 CN 122138836 A Figure 5 Instruction manual illustration 5 / 10 pages 33 CN 122138836 A Figure 6 Instruction manual illustration 6 / 10 pages 34 CN 122138836 A Figure 7 Figure 8 Instruction manual illustration 7 / 10 pages 35 CN 122138836 A Instruction manual illustration 8 / 10 pages 36 CN 122138836 A Figure 9C Instruction manual illustration 9 / 10 pages 37 CN 122138836 A Figure 10 The instruction manual includes attached figures on page 10 / 10, document number 38, CN 122138836 A.

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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